US20030166868A1 - Polypeptide variants - Google Patents

Polypeptide variants Download PDF

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US20030166868A1
US20030166868A1 US09/792,938 US79293801A US2003166868A1 US 20030166868 A1 US20030166868 A1 US 20030166868A1 US 79293801 A US79293801 A US 79293801A US 2003166868 A1 US2003166868 A1 US 2003166868A1
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region
polypeptide
variant
amino acid
binding
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Leonard Presta
Robert Shields
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Genentech Inc
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Genentech Inc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2896—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against molecules with a "CD"-designation, not provided for elsewhere
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6873—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting an immunoglobulin; the antibody being an anti-idiotypic antibody
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/734—Complement-dependent cytotoxicity [CDC]

Definitions

  • the present invention concerns variants of polypeptides comprising an Fc region. More particularly, the present invention concerns Fc region-containing polypeptides that have altered effector function as a consequence of one or more amino acid substitutions in the Fc region of the nonvariant polypeptide.
  • the invention also relates to novel immune complexes and an assay for determining binding of an analyte, such as an Fc region-containing polypeptide, to a receptor.
  • Antibodies are proteins, which exhibit binding specificity to a specific antigen.
  • Native antibodies are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies between the heavy chains of different immunoglobulin isotypes.
  • Each heavy and light chain also has regularly spaced intrachain disulfide bridges.
  • Each heavy chain has at one end a variable domain (V H ) followed by a number of constant domains.
  • V H variable domain
  • Each light chain has a variable domain at one end (V L ) and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light and heavy chain variable domains.
  • variable refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies and are responsible for the binding specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed through the variable domains of antibodies. It is concentrated in three segments called complementarity determining regions (CDRs) both in the light chain and the heavy chain variable domains. The more highly conserved portions of the variable domains are called the framework regions (FR).
  • CDRs complementarity determining regions
  • FR framework regions
  • the variable domains of native heavy and light chains each comprise four FR regions, largely adopting a ⁇ sheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the ⁇ -sheet structure.
  • the CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen binding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest , 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).
  • the constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions.
  • antibodies or immunoglobulins can be assigned to different classes.
  • the heavy chain constant regions that correspond to the different classes of immunoglobulins are called ⁇ , ⁇ , ⁇ , ⁇ , and ⁇ , respectively.
  • human immunoglobulin classes only human IgG1, IgG2, IgG3 and IgM are known to activate complement.
  • FIG. 1 A schematic representation of the native IgG1 structure is shown in FIG. 1, where the various portions of the native antibody molecule are indicated.
  • Papain digestion of antibodies produces two identical antigen binding fragments, called Fab fragments, each with a single antigen binding site, and a residual “Fc” fragment, whose name reflects its ability to crystallize readily.
  • the crystal structure of the human IgG Fc region has been determined (Deisenhofer, Biochemistry 20:2361-2370 (1981)).
  • the Fc region is generated by papain cleavage N-terminal to Cys 226.
  • the Fc region is central to the effector functions of antibodies.
  • effector functions mediated by the antibody Fc region can be divided into two categories: (1) effector functions that operate after the binding of antibody to an antigen (these functions involve the participation of the complement cascade or Fc receptor (FcR)-bearing cells); and (2) effector functions that operate independently of antigen binding (these functions confer persistence in the circulation and the ability to be transferred across cellular barriers by transcytosis).
  • effector functions that operate after the binding of antibody to an antigen (these functions involve the participation of the complement cascade or Fc receptor (FcR)-bearing cells); and (2) effector functions that operate independently of antigen binding (these functions confer persistence in the circulation and the ability to be transferred across cellular barriers by transcytosis).
  • FcR complement cascade or Fc receptor
  • an antibody While binding of an antibody to the requisite antigen has a neutralizing effect that might prevent the binding of a foreign antigen to its endogenous target (e.g. receptor or ligand), binding alone may not remove the foreign antigen.
  • an antibody To be efficient in removing and/or destructing foreign antigens, an antibody should be endowed with both high affinity binding to its antigen, and efficient effector functions.
  • C1q and two serine proteases, C1r and C1s form the complex C1, the first component of the complement dependent cytotoxicity (CDC) pathway.
  • C1q is a hexavalent molecule with a molecular weight of approximately 460,000 and a structure likened to a bouquet of tulips in which six collagenous “stalks” are connected to six globular head regions.
  • Burton and Woof Advances in Immunol . 51:1-84 (1992).
  • C1q To activate the complement cascade, it is necessary for C1q to bind to at least two molecules of IgG1, IgG2, or IgG3 (the consensus is that IgG4 does not activate complement), but only one molecule of IgM, attached to the antigenic target. Ward and Ghetie, Therapeutic Immunology 2:77-94 (1995) at page 80.
  • the residue Pro331 has been implicated in C1q binding by analysis of the ability of human IgG subclasses to carry out complement mediated cell lysis. Mutation of Ser331 to Pro331 in IgG4 conferred the ability to activate complement. (Tao et aL, J. Exp. Med ., 178:661-667 (1993); Brekke et al., Eur. J. lmmunol , 24:2542-47 (1994)).
  • Effector functions can also be mediated by the interaction of the Fc region of an antibody with Fc receptors (FcRs), which are specialized cell surface receptors on hematopoietic cells.
  • Fc receptors belong in the immunoglobulin superfamily, and have been shown to mediate both the removal of antibody-coated pathogens by phagocytosis of immune complexes, and the lysing of erythrocytes and various other cellular targets (e.g. tumor cells) coated with the corresponding antibody, via antibody dependent cell mediated cytotoxicity (ADCC). Van de Winkel and Anderson, J. Leuk. Biol . 49:511-24 (1991).
  • FcRs are defined by their specificity for immunoglobulin isotypes; Fc receptors for IgG antibodies are referred to as Fc ⁇ R, for IgE as Fc ⁇ R, for IgA as Fc ⁇ R and so on.
  • Fc ⁇ RI CD64
  • Fc ⁇ RII CD32
  • Fc ⁇ RIII CD16
  • the three genes encoding the Fc ⁇ RI subclass are clustered in region 1q21.1 of the long arm of chromosome 1; the genes encoding Fc ⁇ RII isoforms (Fc ⁇ RIIA, Fc ⁇ RIIB and Fc ⁇ RIIC) and the two genes encoding Fc ⁇ RIII (Fc ⁇ RIIIA and Fc ⁇ RIIIB) are all clustered in region 1q22.
  • FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et aL, Immunomethods 4:25-34 (1994); and de Haas etal., J. Lab. Clin. Med . 126:330-41 (1995).
  • Fc ⁇ RI binds monomeric IgG with a high affinity
  • Fc ⁇ RII and Fc ⁇ RIII are low-affinity receptors, interacting with complexed or aggregated IgG.
  • the classical method for detecting these low-affinity receptors is by “rosetting” using antibody-coated erythrocytes (EA) sensitized with IgGs.
  • EA antibody-coated erythrocytes
  • Bredius et aL evaluated rosette formation between IgG-sensitized red blood cells and polymorphonuclear leukocytes (PMN) which express Fc ⁇ RIIa and Fc ⁇ RIIIb at their cell-surface. Rosette was defined as three of more EA bound per PMN (Bredius etal.
  • IgG dimers have been formed using anti-light chain monoclonal antibodies (Nagarajan et al, supra and Warmerdam et al., supra) or chemical cross-linking agents (Hogarth et al. Immunomethods 4:17-24 (1994); and Tamm et aL J. Biol Chem . 271(7):3659-3666(1996)). Heat-aggregated immune complexes have also been evaluated for binding to cells expressing FcRs (Tax et al., supra and Tam et al., supra).
  • Pro331 in IgG3 was changed to Ser, and the affinity of this mutant to target cells analyzed. The affinity was found to be six-fold lower than that of unmutated IgG3, indicating the involvement of Pro331 in Fc ⁇ RI binding. Morrison et al., Immunologist , 2:119-124 (1994); and Canfield and Morrison, J. Exp. Med . 173:1483-91 (1991).
  • Glu318 was identified as being involved in binding to Fc ⁇ RII. Ward and Ghetie, Therapeutic Immunology 2:77-94 (1995).
  • the present invention provides a variant of a polypeptide comprising a human IgG Fc region, which variant comprises an amino acid substitution at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
  • the invention further relates to a variant of a polypeptide comprising a human IgG Fc region, which variant binds Fc ⁇ RI, Fc ⁇ RII, Fc ⁇ RIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 322 or amino acid position 329, or both amino acid positions of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
  • the invention provides a method for modifying a polypeptide comprising a human IgG Fc region comprising substituting an amino acid residue at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human lgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
  • the invention also provides a composition comprising the polypeptide variant and a physiologically acceptable carrier or diluent.
  • This composition for potential therapeutic use is sterile and may be lyophilized.
  • the invention provides a method for determining the presence of a protein of interest comprising exposing a sample suspected of containing the protein to the polypeptide variant and determining binding of the polypeptide variant to the sample.
  • the invention provides a method of treating a mammal suffering from a disorder comprising administering to the mammal a therapeutically effective amount of a variant of a polypeptide comprising a human IgG Fc region, which variant binds Fc ⁇ RI, Fc ⁇ RII, Fc ⁇ RIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 329, 331 or 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
  • the invention further provides: isolated nucleic acid encoding the polypeptide variant; a vector comprising the nucleic acid, optionally, operably linked to control sequences recognized by a host cell transformed with the vector; a host cell comprising the vector; a process for producing the polypeptide variant comprising culturing this host cell so that the nucleic acid is expressed and, optionally, recovering the polypeptide variant from the host cell culture (e.g. from the host cell culture medium).
  • the invention also pertains to an immune complex comprising: (a) an Fc region-containing polypeptide; (b) a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide; and (c) a second target molecule comprises at least two binding sites for the first target molecule.
  • the immune complex may be used in an FcR-binding assay, particularly where the FcR has a low affinity for the Fc region-containing polypeptide. Other uses for the immune complex are disclosed herein.
  • the invention provides a method for determining binding of an analyte, such as an Fc region-containing polypeptide, to a receptor (e.g. a low affinity FcR) comprising the following steps performed sequentially: (a) forming a molecular complex between the analyte and a first target molecule, wherein the first target molecule comprises at least two binding sites for the analyte; and (b) determining binding of the molecular complex of step (a) to the receptor (e.g. to a binding domain of the receptor coated on an assay plate).
  • the molecular complex of step (a) further comprises a second target molecule which comprises at least two binding sites for the first target molecule.
  • the invention also relates to an assay kit, such as a kit useful for determining binding of an analyte to a receptor comprising: (a) a first target molecule which comprises at least two binding sites for the analyte; and (b) a second target molecule which comprises at least two binding sites for the first target molecule.
  • FIG. 1 is a schematic representation of a native IgG. Disulfide bonds are represented by heavy lines between CH1 and CL domains and the two CH2 domains. V is variable domain; C is constant domain; L stands for light chain and H stands for heavy chain.
  • FIG. 2 shows C1q binding of wild type (wt) C2B8 antibody; C2B8 antibody with a human IgG2 constant region (IgG2); and mutants K322A, K320A and E318A.
  • FIG. 3 depicts C1q binding of mutants P331A, P329A and K322A.
  • FIGS. 4A and 4B depict the amino acid sequences of E27 anti-IgE antibody light chain (FIG. 4A) and heavy chain (FIG. 4B).
  • FIG. 5 is a schematic diagram of the “immune complex” prepared for use in the FcR assay described in Example 1.
  • the hexamer comprising three anti-IgE antibody molecules (the “Fc region-containing polypeptide”) and three IgE molecules (the “first target molecule”) is shown.
  • IgE has two “binding sites” for the anti-lgE antibody (E27) in the Fc region thereof.
  • Each IgE molecule in the complex is further able to bind two VEGF molecules (“the second target polypeptide”).
  • VEGF has two “binding sites” for IgE.
  • the numbering of the residues in an IgG heavy chain is that of the EU index as in Kabat et al., Sequences of Proteins of Immunological Interest , 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991), expressly incorporated herein by reference.
  • the “EU index as in Kabat” refers to the residue numbering of the human IgG1 EU antibody.
  • Fc region is used to define a C-terminal region of an IgG heavy chain as shown FIG. 1. Although the boundaries of the Fc region of an IgG heavy chain might vary slightly, the human IgG heavy chain Fc region is usually defined to stretch from amino acid residue at position Cys226 to the carboxyl-terminus.
  • Fc region-containing polypeptide refers to a polypeptide, such as an antibody or immunoadhesin (see definitions below), which comprises an Fc region.
  • the Fc region of an IgG comprises two constant domains, CH2 and CH3, as shown in FIG. 1.
  • the “CH2” domain of a human IgG Fc region (also referred to as “C ⁇ 2” domain) usually extends from amino acid 231 to amino acid 340.
  • the CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgG molecule. It has been speculated that the carbohydrate may provide a substitute for the domain-domain pairing and help stabilize the CH2 domain.
  • Hinge region is generally defined as stretching from Glu216 to Pro230 of human IgG1 (Burton, Molec. Immunol . 22:161-206 (1985)) Hinge regions of other IgG isotypes may be aligned with the IgG1 sequence by placing the first and last cysteine residues forming inter-heavy chain S-S bonds in the same positions.
  • C1q is a polypeptide that includes a binding site for the Fc region of an immunoglobulin. C1q together with two serine proteases, C1r and C1s, forms the complex C1, the first component of the complement dependent cytotoxicity (CDC) pathway. Human C1q can be purchased commercially from, e.g. Quidel, San Diego, CA.
  • Fc receptor or “FcR” is used to describe a receptor that binds to the Fc region of an antibody.
  • the preferred FcR is one, which binds an IgG antibody (a gamma receptor) and includes receptors of the Fc ⁇ RI, Fc ⁇ RII, and Fc ⁇ RIII subclasses, including allelic variants and alternatively spliced forms of these receptors.
  • FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al, Immunomethods 4:25-34 (1994); and de Haas et al, J. Lab. Clin. Med . 126:330-41 (1995).
  • FcR FcR
  • FcRn neonatal receptor
  • binding domain refers to the region of a polypeptide that binds to another molecule.
  • the binding domain can comprise a portion of a polypeptide chain thereof (e.g. the ⁇ chain thereof) which is responsible for binding an Fc region.
  • One useful binding domain is the extracellular domain of an FcR ⁇ chain.
  • antibody is used in the broadest sense and specifically covers monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments so long as they exhibit the desired biological activity.
  • Antibody fragments comprise a portion of an intact antibody, generally including the antigen binding or variable region of the intact antibody, so long as they retain at least the CH2 region of an IgG immunoglobulin heavy chain constant domain, comprising amino acid residues 322, 329 and 331, and have the ability, alone or in combination with another antibody fragment, to specifically bind a selected antigen.
  • Examples of antibody fragments include linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.
  • the antibody fragments preferably retain at least part of the hinge and optionally the CH1 region of an IgG heavy chain. More preferably, the antibody fragments retain the entire constant region of an IgG heavy chain, and include an IgG light chain.
  • the term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional (polyclonal) antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen.
  • the modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
  • the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al, Nature 256:495 (1975), or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567).
  • the “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature 352:624-628 (1991) and Marks et al., J. Mol. Biol . 222:581-597 (1991), for example.
  • the monoclonal antibodies herein specifically include “chimeric” antibodies (immunoglobulins) in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al, Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)).
  • chimeric antibodies immunoglobulins in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in
  • “Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin.
  • humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity.
  • donor antibody such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity.
  • Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues.
  • humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance.
  • the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence.
  • the humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.
  • Fc immunoglobulin constant region
  • hypervariable region when used herein refers to the amino acid residues of an antibody which are responsible for antigen-binding.
  • the hypervariable region comprises amino acid residues from a “complementarity determining region” or “CDR” (i.e. residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2) and 95-102 (H3) in the heavy chain variable domain; Kabat et al., Sequences of Proteins of Immunological Interest , 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md.
  • CDR complementarity determining region
  • residues from a “hypervariable loop” i.e. residues 26-32 (L1), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (H1), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain; Chothia and Lesk J. Mol. Biol . 196:901-917 (1987)).
  • “Framework” or “FR” residues are those variable domain residues other than the hypervariable region residues as herein defined.
  • the term “immunoadhesin” designates antibody-like molecules which combine the “binding domains” of a heterologous “adhesin” protein (e.g. a receptor, ligand or enzyme) with an immunoglobulin constant domain.
  • adhesin e.g. a receptor, ligand or enzyme
  • the immunoadhesins comprise a fusion of the adhesin amino acid sequence with the desired binding specificity which is other than the antigen recognition and binding site (antigen combining site) of an antibody (i.e. is “heterologous”) and an immunoglobulin constant domain sequence.
  • ligand binding domain refers to any native cell-surface receptor or any region or derivative thereof retaining at least a qualitative ligand binding ability of a corresponding native receptor.
  • the receptor is from a cell-surface polypeptide having an extracellular domain that is homologous to a member of the immunoglobulin supergenefamily.
  • receptors which are not members of the immunoglobulin supergenefamily but are nonetheless specifically covered by this definition, are receptors for cytokines, and in particular receptors with tyrosine kinase activity (receptor tyrosine kinases), members of the hematopoietin and nerve growth factor receptor superfamilies, and cell adhesion molecules, e.g. (E-, L- and P-) selectins.
  • receptor binding domain is used to designate any native ligand for a receptor, including cell adhesion molecules, or any region or derivative of such native ligand retaining at least a qualitative receptor binding ability of a corresponding native ligand. This definition, among others, specifically includes binding sequences from ligands for the above-mentioned receptors.
  • an “antibody-immunoadhesin chimera” comprises a molecule that combines at least one binding domain of an antibody (as herein defined) with at least one immunoadhesin (as defined in this application).
  • Exemplary antibody-immunoadhesin chimeras are the bispecific CD4-lgG chimeras described in Berg et al, PNAS ( USA ) 88:4723-4727 (1991) and Chamow et al, J. Immunol . 153:4268 (1994).
  • An “isolated” polypeptide is one that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the polypeptide, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes.
  • the polypeptide will be purified (1) to greater than 95% by weight of polypeptide as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain.
  • Isolated polypeptide includes the polypeptide in situ within recombinant cells since at least one component of the polypeptide's natural environment will not be present. Ordinarily, however, isolated polypeptide will be prepared by at least one purification step.
  • Treatment refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented.
  • a “disorder” is any condition that would benefit from treatment with the polypeptide variant. This includes chronic and acute disorders or diseases including those pathological conditions which predispose the mammal to the disorder in question.
  • label when used herein refers to a detectable compound or composition which is conjugated directly or indirectly to the polypeptide.
  • the label may be itself be detectable (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which is detectable.
  • An “isolated” nucleic acid molecule is a nucleic acid molecule that is identified and separated from at least one contaminant nucleic acid molecule with which it is ordinarily associated in the natural source of the polypeptide nucleic acid.
  • An isolated nucleic acid molecule is other than in the form or setting in which it is found in nature. Isolated nucleic acid molecules therefore are distinguished from the nucleic acid molecule as it exists in natural cells.
  • an isolated nucleic acid molecule includes a nucleic acid molecule contained in cells that ordinarily express the polypeptide where, for example, the nucleic acid molecule is in a chromosomal location different from that of natural cells.
  • control sequences refers to DNA sequences necessary for the expression of an operably linked coding sequence in a particular host organism.
  • the control sequences that are suitable for prokaryotes include a promoter, optionally an operator sequence, and a ribosome binding site.
  • Eukaryotic cells are known to utilize promoters, polyadenylation signals, and enhancers.
  • Nucleic acid is “operably linked” when it is placed into a functional relationship with another nucleic acid sequence.
  • DNA for a presequence or secretory leader is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide;
  • a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or
  • a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation.
  • “operably linked” means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading phase. However, enhancers do not have to be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, the synthetic oligonucleotide adaptors or linkers are used in accordance with conventional practice.
  • the expressions “cell,” “cell line,” and “cell culture” are used interchangeably and all such designations include progeny.
  • the words “transformants” and “transformed cells” include the primary subject cell and cultures derived therefrom without regard for the number of transfers. It is also understood that all progeny may not be precisely identical in DNA content, due to deliberate or inadvertent mutations. Mutant progeny that have the same function or biological activity as screened for in the originally transformed cell are included. Where distinct designations are intended, it will be clear from the context.
  • molecular complex when used herein refers to the relatively stable structure which forms when two or more heterologous molecules (e.g. polypeptides) bind (preferably noncovalently) to one another.
  • the preferred molecular complex herein is an immune complex.
  • Immuno complex refers to the relatively stable structure which forms when at least one target molecule and at least one heterologous Fc region-containing polypeptide bind to one another forming a larger molecular weight complex.
  • immune complexes are antigen-antibody aggregates and target molecule-immunoadhesin aggregates.
  • immuno complex as used herein, unless indicated otherwise, refers to an ex vivo complex (i.e. other than the form or setting in which it may be found in nature). However, the immune complex may be administered to a mammal, e.g. to evaluate clearance of the immune complex in the mammal.
  • target molecule refers to a molecule, usually a polypeptide, which is capable of being bound by a heterologous molecule and has one or more binding sites for the heterologous molecule.
  • binding site refers to a region of a molecule to which another molecule can bind.
  • the “first target molecule” herein comprises at least two distinct binding sites (for example, two to five separate binding sites) for an analyte (e.g. an Fc region-containing polypeptide) such that at least two analyte molecules can bind to the first target molecule.
  • the two or more binding sites are identical (e.g. having the same amino acid sequence, where the target molecule is a polypeptide).
  • the first target molecule was IgE and had two separate binding sites in the Fc region thereof to which the Fc region-containing polypeptide (an anti-IgE antibody, E27) could bind.
  • Other first target molecules include dimers of substantially identical monomors (e.g. neurotrophins, IL8 and VEGF) or are polypeptides comprising two or more substantially identical polypeptide chains (e.g. antibodies or immunoadhesins).
  • the “second target molecule” comprises at least two distinct binding sites (for example, two to five separate binding sites) for the first target molecule such that at least two first target molecules can bind to the second target molecule.
  • the two or more binding sites are identical (e.g.
  • the second target molecule was VEGF, which has a pair of distinct binding sites to which the variable domain of the IgE antibody could bind.
  • Other second target molecules are contemplated, e.g. other dimers of substantially identical monomers (e.g. neurotrophins or IL8) or polypeptides comprising two or more substantially identical domains (e.g. antibodies or immunoadhesins).
  • an “analyte” is a substance that is to be analyzed.
  • the preferred analyte is an Fc region-containing polypeptide that is to be analyzed for its ability to bind to an Fc receptor.
  • a “receptor” is a polypeptide capable of binding at least one ligand.
  • the preferred receptor is a cell-surface receptor having an extracellular ligand-binding domain and, optionally, other domains (e.g. transmembrane domain, intracellular domain and/or membrane anchor).
  • the receptor to be evaluated in the assay described herein may be an intact receptor or a fragment or derivative thereof (e.g. a fusion protein comprising the binding domain of the receptor fused to one or more heterologous polypeptides).
  • the receptor to be evaluated for its binding properties may be present in a cell or isolated and optionally coated on an assay plate or some other solid phase.
  • low affinity receptor denotes a receptor that has a weak binding affinity for a ligand of interest, e.g. having a binding constant of about 50 nM or worse affinity.
  • exemplary low affinity receptors include Fc ⁇ RII and Fc ⁇ RIII as well as adhesion molecules, such as integrins.
  • the invention herein relates to a method for making a polypeptide variant.
  • the “starting” or “nonvariant” polypeptide is prepared using techniques available in the art for generating polypeptides comprising an Fc region.
  • the polypeptide is an antibody and exemplary methods for generating antibodies are described in more detail in the following sections.
  • the polypeptide may, however, be any other polypeptide comprising an Fc region, e.g. an immunoadhesin. Methods for making immunoadhesins are elaborated in more detail hereinbelow.
  • the starting polypeptide of particular interest herein is usually one that binds to C1q and displays complement dependent cytotoxicity (CDC).
  • CDC complement dependent cytotoxicity
  • the amino acid substitutions described herein will generally serve to alter the ability of the starting polypeptide to bind to C1q and/or modify its complement dependent cytotoxicity function, e.g. to reduce and preferably abolish these effector functions.
  • polypeptides comprising substitutions at one or more of the described positions with improved effector functions are contemplated herein.
  • the starting polypeptide may be unable to bind C1q and/or mediate CDC and may be modified according to the teachings herein such that it acquires these effector functions.
  • polypeptides with pre-existing C1q binding activity, optionally further having the ability to mediate CDC may be modified such that one or both of these activities are enhanced.
  • one or more amino acid alterations are introduced in the Fc region of the starting polypeptide.
  • the amino acid positions to be modified are selected from heavy chain positions 329, 331 and 322, where the numbering of the residues in an IgG heavy chain is that of the EU index as in Kabat et al, Sequences of Proteins of Immunological Interest , 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md (1991).
  • the Fc region is preferably from a human IgG and most preferably a human IgG1.
  • the human IgG1 Fc region may be a human A or non-A allotype.
  • Proline is conserved at position 329 in human IgG's. This residue is preferably replaced with alanine, however substitution with any other amino acid is contemplated, e.g., serine, threonine, asparagine, glycine or valine. Proline is conserved at position 331 in human IgG1, IgG2 and IgG3, but not IgG4 (which has a serine residue at position 331). Residue 331 is preferably replaced by alanine or another amino acid, e.g. serine (for IgG regions other than IgG4), glycine or valine.
  • Lysine 322 is conserved in human IgGs, and this residue is preferably replaced by an alanine residue, but substitution with any other amino acid residue is contemplated, e.g. serine, threonine, glycine or valine.
  • only one of the three above-identified positions is altered in order to generate the polypeptide variant.
  • residue 329 or only residue 322 is altered if this is the case.
  • two or all of the three identified positions are modified.
  • a variant may be generated in which the native amino acid residue at position 329 of the human heavy chain constant region is substituted with another amino acid, optionally in combination with a substitution of the amino acid residue at position 331 and/or substitution of the amino acid residue at position 322.
  • the native amino acid residue at position 331 and the native amino acid residue at position 322 of the human IgG Fc region may both be substituted with another amino acid residue.
  • DNA encoding amino acid sequence variant of the starting polypeptide is prepared by a variety of methods known in the art. These methods include, but are not limited to, preparation by site-directed (or oligonucleotide-mediated) mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlier prepared DNA encoding the polypeptide
  • Site-directed mutagenesis is a preferred method for preparing substitution variants. This technique is well known in the art (see, e.g.,Carter et al. Nucleic Acids Res . 13:4431-4443 (1985) and Kunkel et al, Proc. Natl. Acad.Sci.USA 82:488 (1987)). Briefly, in carrying out site-directed mutagenesis of DNA, the starting DNA is altered by first hybridizing an oligonucleotide encoding the desired mutation to a single strand of such starting DNA.
  • a DNA polymerase is used to synthesize an entire second strand, using the hybridized oligonucleotide as a primer, and using the single strand of the starting DNA as a template.
  • the oligonucleotide encoding the desired mutation is incorporated in the resulting double-stranded DNA.
  • PCR mutagenesis is also suitable for making amino acid sequence variants of the starting polypeptide. See Higuchi, in PCR Protocols, pp . 177-183 (Academic Press, 1990); and Vallette et al., Nuc. Acids Res . 17:723-733 (1989). Briefly, when small amounts of template DNA are used as starting material in a PCR, primers that differ slightly in sequence from the corresponding region in a template DNA can be used to generate relatively large quantities of a specific DNA fragment that differs from the template sequence only at the positions where the primers differ from the template.
  • the starting material is the plasmid (or other vector) comprising the starting polypeptide DNA to be mutated.
  • the codon(s) in the starting DNA to be mutated are identified. There must be a unique restriction endonuclease site on each side of the identified mutation site(s). If no such restriction sites exist, they may be generated using the above-described oligonucleotide-mediated mutagenesis method to introduce them at appropriate locations in the starting polypeptide DNA.
  • the plasmid DNA is cut at these sites to linearize it.
  • a double-stranded oligonucleotide encoding the sequence of the DNA between the restriction sites but containing the desired mutation(s) is synthesized using standard procedures, wherein the two strands of the oligonucleotide are synthesized separately and then hybridized together using standard techniques.
  • This double-stranded oligonucleotide is referred to as the cassette.
  • This cassette is designed to have 5′ and 3′ ends that are compatible with the ends of the linearized plasmid, such that it can be directly ligated to the plasmid.
  • This plasmid now contains the mutated DNA sequence.
  • polypeptide variant(s) so prepared may be subjected to further modifications, oftentimes depending on the intended use of the polypeptide. Such modifications may involve further alteration of the amino acid sequence, fusion to heterologous polypeptide(s) and/or covalent modifications. For example, it may be useful to combine the above amino acid substitutions with one or more further amino acid substitutions that reduce or ablate FcR binding.
  • the native amino acid residues at any one or more of heavy chain positions 233-238, 318 or 331 may be replaced with non-native residue(s), e.g. alanine.
  • any cysteine residue not involved in maintaining the proper conformation of the polypeptide variant also may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant cross linking.
  • Another type of amino acid substitution serves to alter the glycosylation pattern of the polypeptide. This may be achieved by deleting one or more carbohydrate moieties found in the polypeptide, and/or adding one or more glycosylation sites that are not present in the polypeptide.
  • Glycosylation of polypeptides is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue.
  • the tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain.
  • O-linked glycosylation refers to the attachment of one of the sugars N-aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used.
  • Addition of glycosylation sites to the polypeptide is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites).
  • the alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original polypeptide (for O-linked glycosylation sites).
  • An exemplary glycosylation variant has an amino acid substitution of residue Asn 297 of the heavy chain.
  • the polypeptide variant may be subjected to one or more assays to evaluate any change in biological activity compared to the starting polypeptide. For example, the ability of the variant to bind C1q and mediate complement dependent cytotoxicity (CDC) may be assessed.
  • CDC mediate complement dependent cytotoxicity
  • a C1q binding ELISA may be performed. Briefly, assay plates may be coated overnight at 4° C. with polypeptide variant or starting polypeptide (control) in coating buffer. The plates may then be washed and blocked. Following washing, an aliquot of human C1q may be added to each well and incubated for 2 hrs at room temperature. Following a further wash, 100 ⁇ l of a sheep anti-complement C1q peroxidase conjugated antibody may be added to each well and incubated for 1 hour at room temperature. The plate may again be washed with wash buffer and 100 ⁇ l of substrate buffer containing OPD ( O-phenylenediamine dihydrochloride (Sigma)) may be added to each well. The oxidation reaction, observed by the appearance of a yellow color, may be allowed to proceed for 30 minutes and stopped by the addition of 100 ⁇ l of 4.5 N H 2 SO 4 . The absorbance may then read at (492-405) nm.
  • OPD O-phenylenediamine dihydr
  • the preferred polypeptide variant is one that displays a “significant reduction in C1q binding” in this assay. This means that about 100 ⁇ g/ml of the polypeptide variant displays about 50 fold or more reduction in C1q binding compared to 100 ⁇ g/ml of a control antibody having a nonmutated IgG1 Fc region. In the most preferred embodiment, the polypeptide variant “does not bind C1q”, i.e. 100 ⁇ g/ml of the polypeptide variant displays about 100 fold or more reduction in C1q binding compared to 100 ⁇ g/ml of the control antibody.
  • a complement dependent cytotoxicity (CDC) assay may be performed, e.g. as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996). Briefly, various concentrations of the polypeptide variant and human complement may be diluted with buffer. Cells which express the antigen to which the polypeptide variant binds may be diluted to a density of ⁇ 1 ⁇ 10 6 cells/ ml. Mixtures of polypeptide variant, diluted human complement and cells expressing the antigen may be added to a flat bottom tissue culture 96 well plate and allowed to incubate for 2 hrs at 37°C. and 5% CO 2 to facilitate complement mediated cell lysis.
  • CDC complement dependent cytotoxicity
  • alamar blue (Accumed International) may then be added to each well and incubated overnight at 37°C.
  • the absorbance is measured using a 96-well fluorometer with excitation at 530 nm and emission at 590 nm.
  • the results may be expressed in relative fluorescence units (RFU).
  • the sample concentrations may be computed from a standard curve and the percent activity as compared to nonvariant polypeptide is reported for the polypeptide variant of interest.
  • the preferred variant “does not activate complement”.
  • 0.6 ⁇ /ml of the polypeptide variant displays about 0-10% CDC activity in this assay compared to a 0.6 ⁇ /ml of a control antibody having a nonmutated IgG1 Fc region.
  • the variant does not appear to have any CDC activity in the above CDC assay.
  • the polypeptide variant essentially retains the ability to bind antigen compared to the nonvariant polypeptide, i.e. the binding capability is no worse than about 20 fold, e.g. no worse than about 5 fold of that of the nonvariant polypeptide.
  • the binding capability of the polypeptide variant may be determined using techniques such as fluorescence activated cell sorting (FACS) analysis or radioimmunoprecipitation (RIA), for example.
  • the ability of the polypeptide variant to bind an FcR may also be evaluated.
  • the FcR is a high affinity Fc receptor, such as Fc ⁇ RI or FcRn
  • binding can be measured by titrating monomeric polypeptide variant and measuring bound polypeptide variant using an antibody which specifically binds to the polypeptide variant in a standard ELISA format (see Example 2 below).
  • Another FcR binding assay for low affinity FcRs is elaborated in more detail in the following section.
  • the variant retains the ability to bind one or more FcRs, e.g. the ability of the polypeptide variant to bind Fc ⁇ RI, Fc ⁇ RII, Fc ⁇ RIII and/or FcRn is no more than about 20 fold reduced, preferably no more than about 10 fold reduced, and most preferably no more than about two fold reduced compared to the starting polypeptide as determined in the Fc ⁇ RI or FcRn assays of Example 2 or the Fc ⁇ RII or Fc ⁇ RIII assays described in the following section.
  • FcRs e.g. the ability of the polypeptide variant to bind Fc ⁇ RI, Fc ⁇ RII, Fc ⁇ RIII and/or FcRn is no more than about 20 fold reduced, preferably no more than about 10 fold reduced, and most preferably no more than about two fold reduced compared to the starting polypeptide as determined in the Fc ⁇ RI or FcRn assays of Example 2 or the Fc ⁇ RII or Fc ⁇ RIII assays described
  • a receptor binding assay has been developed herein which is particularly useful for determining binding of an analyte of interest to a receptor where the affinity of the analyte for the receptor is relatively weak, e.g. in the micromolar range as is the case for Fc ⁇ RIIa, Fc ⁇ RIIb, Fc ⁇ RIIIa and Fc ⁇ RIIIb.
  • the method involves the formation of a molecular complex that has an improved avidity for the receptor of interest compared to the noncomplexed analyte.
  • the preferred molecular complex is an immune complex comprising: (a) an Fc region-containing polypeptide (such as an antibody or an immunoadhesin); (b) a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide; and (c) a second target molecule which comprises at least two binding sites for the first target molecule.
  • an Fc region-containing polypeptide such as an antibody or an immunoadhesin
  • a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide
  • a second target molecule which comprises at least two binding sites for the first target molecule.
  • the Fc region-containing polypeptide is an anti-IgE antibody, such as the E27 antibody (FIGS. 4 A- 4 B).
  • E27 when mixed with human IgE at an 1:1 molar ratio, forms a stable hexamer consisting of three E27 molecules and three IgE molecules.
  • the “first target molecule” is a chimeric form of IgE in which the Fab portion of an anti-VEGF antibody is fused to the human IgE Fc portion and the “second target molecule” is the antigen to which the Fab binds (i.e. VEGF). Each molecule of IgE binds two molecules of VEGF.
  • VEGF also binds two molecules of IgE per molecule of VEGF.
  • recombinant human VEGF was added at a 2:1 molar ratio to IgE:E27 hexamers, the hexamers were linked into larger molecular weight complexes via the IgE:VEGF interaction (FIG. 5).
  • the Fc region of the anti-IgE antibody of the resultant immune complex binds to FcR with higher avidity than either uncomplexed anti-IgE or anti-IgE:lgE hexamers.
  • Fc region-containing polypeptide:first target molecule combination examples include an immunoadhesin:ligand combination such as VEGF receptor (KDR)-immunoadhesin: VEGF and a full-length bispecific antibody (bsAb):first target molecule.
  • an Fc region-containing polypeptide:first target molecule:second target molecule combination include a nonblocking antibody:soluble receptor:ligand combination such as anti-Trk antibody:soluble Trk receptor:neurotrophin (Urfer et al. J. Biol Chem . 273(10):5829-5840 (1998)).
  • the immune complexes described above have further uses including evaluation of Fc region-containing polypeptide function and immune complex clearance in vivo.
  • the immune complex may be administered to a mammal (e.g. in a pre-clinical animal study) and evaluated for its half-life etc.
  • a polypeptide comprising at least the binding domain of the receptor of interest may be coated on solid phase, such as an assay plate.
  • the binding domain of the receptor alone or a receptor-fusion protein may be coated on the plate using standard procedures.
  • receptor-fusion proteins include receptor-glutathione S-transferase (GST) fusion protein, receptor-chitin binding domain fusion protein, receptor-hexaHis tag fusion protein (coated on glutathione, chitin, and nickel coated plates, respectively).
  • a capture molecule may be coated on the assay plate and used to bind the receptor-fusion protein via the non-receptor portion of the fusion protein.
  • examples include anti-hexaHis F(ab′) 2 coated on the assay plate used to capture receptor-hexaHis tail fusion or anti-GST antibody coated on the assay plate used to capture a receptor-GST fusion.
  • binding to cells expressing at least the binding domain of the receptor may be evaluated.
  • the cells may be naturally occurring hematopoietic cells that express the FcR of interest or may be transformed with nucleic acid encoding the FcR or a binding domain thereof such that the binding domain is expressed at the surface of the cell to be tested.
  • the immune complex described hereinabove is added to the receptor-coated plates and incubated for a sufficient period of time such that the analyte binds to the receptor. Plates may then be washed to remove unbound complexes, and binding of the analyte may be detected according to known methods. For example, binding may be detected using a reagent (e.g. an antibody or fragment thereof) which binds specifically to the analyte, and which is optionally conjugated with a detectable label (detectable labels and methods for conjugating them to polypeptides are described below in the section entitled “Non-Therapeutic Uses for the Polypeptide Variant”).
  • a reagent e.g. an antibody or fragment thereof
  • the reagents can be provided in an assay kit, i.e., a packaged combination of reagents, for combination with the analyte in assaying the ability of the analyte to bind to a receptor of interest.
  • the components of the kit will generally be provided in predetermined ratios.
  • the kit may provide the first target molecule and/or the second target molecule, optionally complexed together.
  • the kit may further include assay plates coated with the receptor or a binding domain thereof (e.g. the extracellular domain of the ⁇ subunit of an FcR).
  • kits such as an antibody that binds specifically to the analyte to be assayed, labeled directly or indirectly with an enzymatic label
  • the detectable label is an enzyme
  • the kit will include substrates and cofactors required by the enzyme (e.g. a substrate precursor which provides the detectable chromophore or fluorophore).
  • other additives may be included such as stabilizers, buffers (e.g. assay and/or wash lysis buffer) and the like.
  • the relative amounts of the various reagents may be varied widely to provide for concentrations in solution of the reagents that substantially optimize the sensitivity of the assay.
  • the reagents may be provided as dry powders, usually lyophilized, including excipients that on dissolution will provide a reagent solution having the appropriate concentration.
  • the kit also suitably includes instructions for carrying out the assay.
  • the Fc region-containing polypeptide which is modified according to the teachings herein is an antibody. Techniques for producing antibodies follow:
  • the polypeptide is an antibody
  • it is directed against an antigen of interest.
  • the antigen is a biologically important polypeptide and administration of the antibody to a mammal suffering from a disease or disorder can result in a therapeutic benefit in that mammal.
  • antibodies directed against nonpolypeptide antigens are also contemplated.
  • the antigen is a polypeptide, it may be a transmembrane molecule (e.g. receptor) or ligand such as a growth factor.
  • exemplary antigens include molecules such as renin; a growth hormone, including human growth hormone and bovine growth hormone; growth hormone releasing factor; parathyroid hormone; thyroid stimulating hormone; lipoproteins; alpha-1-antitrypsin; insulin A-chain; insulin B-chain; proinsulin; follicle stimulating hormone; calcitonin; luteinizing hormone; glucagon; clofting factors such as factor VIIIC, factor IX, tissue factor, and von Willebrands factor; anti-clofting factors such as Protein C; atrial natriuretic factor; lung surfactant; a plasminogen activator, such as urokinase or human urine or tissue-type plasminogen activator (t-PA); bombesin; thrombin; hemopoietic growth factor; tumor necrosis factor-alpha
  • Preferred molecular targets for antibodies encompassed by the present invention include CD proteins such as CD3, CD4, CD8, CD19, CD20 and CD34; members of the ErbB receptor family such as the EGF receptor, HER2, HER3 or HER4 receptor; cell adhesion molecules such as LFA-1, Mac1, p150.95, VLA-4, ICAM-1, VCAM and ⁇ v/ ⁇ 3 integrin including either ⁇ or ⁇ subunits thereof (e.g. anti-CD11 a , anti-CD18 or anti-CD11 b antibodies); growth factors such as VEGF; IgE; blood group antigens; flk2/flt3 receptor; obesity (OB) receptor; mpl receptor; CTLA-4; protein C etc.
  • CD proteins such as CD3, CD4, CD8, CD19, CD20 and CD34
  • members of the ErbB receptor family such as the EGF receptor, HER2, HER3 or HER4 receptor
  • cell adhesion molecules such as LFA-1, Mac1, p150.95
  • Soluble antigens or fragments thereof, optionally conjugated to other molecules, can be used as immunogens for generating antibodies.
  • immunogens for transmembrane molecules, such as receptors, fragments of these (e.g. the extracellular domain of a receptor) can be used as the immunogen.
  • transmembrane molecules such as receptors
  • fragments of these e.g. the extracellular domain of a receptor
  • cells expressing the transmembrane molecule can be used as the immunogen.
  • Such cells can be derived from a natural source (e.g. cancer cell lines) or may be cells which have been transformed by recombinant techniques to express the transmembrane molecule.
  • Other antigens and forms thereof useful for preparing antibodies will be apparent to those in the art.
  • Polyclonal antibodies are preferably raised in animals by multiple subcutaneous (sc) or intraperitoneal (ip) injections of the relevant antigen and an adjuvant. It may be useful to conjugate the relevant antigen to a protein that is immunogenic in the species to be immunized, e.g., keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor using a bifunctional or derivatizing agent, for example, maleimidobenzoyl sulfosuccinimide ester (conjugation through cysteine residues), N-hydroxysuccinimide (through lysine residues), glutaraldehyde, succinic anhydride, SOCl 2 , or R 1 N ⁇ C ⁇ NR, where R and R 1 are different alkyl groups.
  • a protein that is immunogenic in the species to be immunized e.g., keyhole limpet hemocyanin, serum albumin, bovine thy
  • Animals are immunized against the antigen, immunogenic conjugates, or derivatives by combining, e.g., 100 ⁇ g or 5 ⁇ g of the protein or conjugate (for rabbits or mice, respectively) with 3 volumes of Freund's complete adjuvant and injecting the solution intradermally at multiple sites.
  • the animals are boosted with 1/5 to 1/10 the original amount of peptide or conjugate in Freund's complete adjuvant by subcutaneous injection at multiple sites.
  • Seven to 14 days later the animals are bled and the serum is assayed for antibody titer. Animals are boosted until the titer plateaus.
  • the animal is boosted with the conjugate of the same antigen, but conjugated to a different protein and/or through a different cross-linking reagent.
  • Conjugates also can be made in recombinant cell culture as protein fusions.
  • aggregating agents such as alum are suitably used to enhance the immune response.
  • Monoclonal antibodies may be made using the hybridoma method first described by Kohler et al, Nature, 256:495 (1975), or may be made by recombinant DNA methods (U.S. Pat. No. 4,816,567).
  • a mouse or other appropriate host animal such as a hamster or macaque monkey
  • lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization.
  • lymphocytes may be immunized in vitro. Lymphocytes then are fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice , pp.59-103 (Academic Press, 1986)).
  • the hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells.
  • a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells.
  • the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells.
  • Preferred myeloma cells are those that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium.
  • preferred myeloma cell lines are murine myeloma lines, such as those derived from MOPC-21 and MPC-11 mouse tumors available from the Salk Institute Cell Distribution Center, San Diego, Calif. USA, and SP-2 or X63-Ag8-653 cells available from the American Type Culture Collection, Rockville, Md. USA.
  • Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol ., 133:3001 (1984); Brodeur et al, Monoclonal Antibody Production Techniques and Applications , pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).
  • Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against the antigen.
  • the binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).
  • RIA radioimmunoassay
  • ELISA enzyme-linked immunoabsorbent assay
  • the clones may be subcloned by limiting dilution procedures and grown by standard methods (Goding, Monoclonal Antibodies: Principles and Practice , pp.59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium.
  • the hybridoma cells may be grown in vivo as ascites tumors in an animal.
  • the monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.
  • DNA encoding the monoclonal antibodies is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the monoclonal antibodies).
  • the hybridoma cells serve as a preferred source of such DNA.
  • the DNA may be placed into expression vectors, which are then transfected into host cells such as E. coi cells, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. Recombinant production of antibodies will be described in more detail below.
  • antibodies or antibody fragments can be isolated from antibody phage libraries generated using the techniques described in McCafferty et al, Nature , 348:552-554 1990). Clackson et al., Nature , 352:624-628 (1991) and Marks et al, J. Mol. Biol ., 222:581-597 (1991) describe the isolation of murine and human antibodies, respectively, using phage libraries.
  • the DNA also may be modified, for example, by substituting the coding sequence for human heavy- and light-chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567; Morrison, et al., Proc. Natl Acad. Sci. USA , 81:6851 (1984)), or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide.
  • non-immunoglobulin polypeptides are substituted for the constant domains of an antibody, or they are substituted for the variable domains of one antigen-combining site of an antibody to create a chimeric bivalent antibody comprising one antigen-combining site having specificity for an antigen and another antigen-combining site having specificity for a different antigen.
  • a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as “import” residues, which are typically taken from an “import” variable domain. Humanization can be essentially performed following the method of Winter and co-workers (Jones et al., Nature, 321:522-525 (1986); Riechmann et al, Nature , 332:323-327 (1988); Verhoeyen et al, Science , 239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Accordingly, such “humanized” antibodies are chimeric antibodies (U.S.
  • humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
  • variable domains both light and heavy
  • the choice of human variable domains, both light and heavy, to be used in making the humanized antibodies is very important to reduce antigenicity.
  • the sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable-domain sequences.
  • the human sequence which is closest to that of the rodent is then accepted as the human framework (FR) for the humanized antibody (Sims et al, J. Immunol ., 151:2296 (1993); Chothia et al., J. Mol Biol , 196:901 (1987)).
  • Another method uses a particular framework derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains.
  • the same framework may be used for several different humanized antibodies (Carter et al., Proc. Natl. Acad. Sci. USA , 89:4285 (1992); Presta et al., J. Immnol ., 151:2623 (1993)).
  • humanized antibodies are prepared by a process of analysis of the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences.
  • Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art.
  • Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen.
  • FR residues can be selected and combined from the recipient and import sequences so that the desired antibody characteristic, such as increased affinity for the target antigen(s), is achieved.
  • the CDR residues are directly and most substantially involved in influencing antigen binding.
  • transgenic animals e.g., mice
  • transgenic animals e.g., mice
  • J H antibody heavy-chain joining region
  • Human antibodies can also be derived from phage-display libraries (Hoogenboom et al., J. MoL Biol ., 227:381 (1991); Marks et al., J. Mol. Biol , 222:581-597 (1991); Vaughan et al. Nature Biotech 14:309 (1996)).
  • Multispecific antibodies have binding specificities for at least two different antigens. While such molecules normally will only bind two antigens (i.e. bispecific antibodies, BsAbs), antibodies with additional specificities such as trispecific antibodies are encompassed by this expression when used herein.
  • BsAbs include those with one arm directed against a tumor cell antigen and the other arm directed against a cytotoxic trigger molecule such as anti-Fc ⁇ Rl/anti-CD15, anti-p185 HER2 /Fc ⁇ RIII (CD16), anti-CD3/anti-malignant B-cell (1D10), anti-CD3/anti-p185 HER2 , anti-CD3/anti-p97, anti-CD3/anti-renal cell carcinoma, anti-CD3/anti-OVCAR-3, anti-CD3/L-D1 (anti-colon carcinoma), anti-CD3/anti-melanocyte stimulating hormone analog, anti-EGF receptor/anti-CD3, anti-CD3/anti-CAMA1, anti-CD3/anti-CD19, anti-CD3/MoV18, anti-neural cell ahesion molecule (NCAM)/anti-CD3, anti-folate binding protein (FBP)/anti-CD3, anti-pan carcinoma associated antigen (AMOC-31)/anti-CD3; BsAbs with one arm which bind
  • BsAbs for use in therapy of infectious diseases such as anti-CD3/anti-herpes simplex virus (HSV), anti-T-cell receptor:CD3 complex/anti-influenza, anti-Fc ⁇ R/anti-HIV; BsAbs for tumor detection in vitro or in vivo such as anti-CEA/anti-EOTUBE, anti-CEA/anti-DPTA, anti-p185 HER2 /anti-hapten; BsAbs as vaccine adjuvants; and BsAbs as diagnostic tools such as anti-rabbit IgG/anti-ferritin, anti-horse radish peroxidase (HRP)/anti-hormone, anti-somatostatin/anti-substance P, anti-HRP/anti-FITC, anti-CEA/anti- ⁇ -galactosidase.
  • HRP anti-horse radish peroxidase
  • HRP anti-somatostatin/anti-substance P
  • trispecific antibodies examples include anti-CD3/anti-CD4/anti-CD37, anti-CD3/anti-CD5/anti-CD37 and anti-CD3/anti-CD8/anti-CD37.
  • Bispecific antibodies can be prepared as full length antibodies or antibody fragments (e.g. F(ab′) 2 bispecific antibodies).
  • antibody variable domains with the desired binding specificities are fused to immunoglobulin constant domain sequences.
  • the fusion preferably is with an immunoglobulin heavy chain constant domain, comprising at least part of the hinge, CH2, and CH3 regions. It is preferred to have the first heavy-chain constant region (CH1) containing the site necessary for light chain binding, present in at least one of the fusions.
  • DNAs encoding the immunoglobulin heavy chain fusions and, if desired, the immunoglobulin light chain are inserted into separate expression vectors, and are co-transfected into a suitable host organism.
  • the bispecific antibodies are composed of a hybrid immunoglobulin heavy chain with a first binding specificity in one arm, and a hybrid immunoglobulin heavy chain-light chain pair (providing a second binding specificity) in the other arm. It was found that this asymmetric structure facilitates the separation of the desired bispecific compound from unwanted immunoglobulin chain combinations, as the presence of an immunoglobulin light chain in only one half of the bispecific molecule provides for a facile way of separation. This approach is disclosed in WO 94/04690. For further details of generating bispecific antibodies see, for example, Suresh et al., Methods in Enzymology , 121:210 (1986).
  • the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers which are recovered from recombinant cell culture.
  • the preferred interface comprises at least a part of the C H 3 domain of an antibody constant domain.
  • one or more small amino acid side chains from the interface of the first antibody molecule are replaced with larger side chains (e.g. tyrosine or tryptophan).
  • Compensatory “cavities” of identical or similar size to the large side chain(s) are created on the interface of the second antibody molecule by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine). This provides a mechanism for increasing the yield of the heterodimer over other unwanted end-products such as homodimers.
  • Bispecific antibodies include cross-linked or “heteroconjugate” antibodies.
  • one of the antibodies in the heteroconjugate can be coupled to avidin, the other to biotin.
  • Such antibodies have, for example, been proposed to target immune system cells to unwanted cells (U.S. Pat. No. 4,676,980), and for treatment of HIV infection (WO 91/00360, WO 921200373, and EP 03089).
  • Heteroconjugate antibodies may be made using any convenient cross-linking methods. Suitable cross-linking agents are well known in the art, and are disclosed in U.S. Pat. No. 4,676,980, along with a number of cross-linking techniques.
  • Antibodies with more than two valencies are contemplated.
  • trispecific antibodies can be prepared. Tutt et al J. Immunol . 147:60 (1991).
  • polypeptide of interest herein is preferably an antibody
  • Fc region-containing polypeptides which can be modified according to the methods described herein are contemplated.
  • An example of such a molecule is an immunoadhesin.
  • the simplest and most straightforward immunoadhesin design combines the binding domain(s) of the adhesin (e.g. the extracellular domain (ECD) of a receptor) with the Fc region of an immunoglobulin heavy chain.
  • ECD extracellular domain
  • nucleic acid encoding the binding domain of the adhesin will be fused C-terminally to nucleic acid encoding the N-terminus of an immunoglobulin constant domain sequence, however N-terminal fusions are also possible.
  • the encoded chimeric polypeptide will retain at least functionally active hinge, C H 2 and C H 3 domains of the constant region of an immunoglobulin heavy chain. Fusions are also made to the C-terminus of the Fc portion of a constant domain, or immediately N-terminal to the C H 1 of the heavy chain or the corresponding region of the light chain.
  • the precise site at which the fusion is made is not critical; particular sites are well known and may be selected in order to optimize the biological activity, secretion, or binding characteristics of the immunoadhesin.
  • the adhesin sequence is fused to the N-terminus of the Fc region of immunoglobulin G 1 (IgG 1 ). It is possible to fuse the entire heavy chain constant region to the adhesin sequence. However, more preferably, a sequence beginning in the hinge region just upstream of the papain cleavage site which defines IgG Fc chemically (i.e. residue 216, taking the first residue of heavy chain constant region to be 114), or analogous sites of other immunoglobulins is used in the fusion.
  • the adhesin amino acid sequence is fused to (a) the hinge region and C H 2 and C H 3 or (b) the C H 1, hinge, C H 2 and C H 3 domains, of an IgG heavy chain.
  • the immunoadhesins are assembled as multimers, and particularly as heterodimers or heterotetramers.
  • these assembled immunoglobulins will have known unit structures.
  • a basic four chain structural unit is the form in which IgG, lgD, and IgE exist.
  • a four chain unit is repeated in the higher molecular weight immunoglobulins; IgM generally exists as a pentamer of four basic units held together by disulfide bonds.
  • IgA globulin, and occasionally IgG globulin may also exist in multimeric form in serum. In the case of multimer, each of the four units may be the same or different.
  • each A represents identical or different adhesin amino acid sequences
  • V L is an immunoglobulin light chain variable domain
  • V H is an immunoglobulin heavy chain variable domain
  • C L is an immunoglobulin light chain constant domain
  • C H is an immunoglobulin heavy chain constant domain
  • n is an integer greater than 1;
  • Y designates the residue of a covalent cross-linking agent.
  • the adhesin sequences can be inserted between immunoglobulin heavy chain and light chain sequences, such that an immunoglobulin comprising a chimeric heavy chain is obtained.
  • the adhesin sequences are fused to the 3′ end of an immunoglobulin heavy chain in each arm of an immunoglobulin, either between the hinge and the C H 2 domain, or between the C H 2 and C H 3 domains. Similar constructs have been reported by Hoogenboom, et al., Mol. Immunol . 28:1027-1037 (1991).
  • an immunoglobulin light chain might be present either covalently associated to an adhesin-immunoglobulin heavy chain fusion polypeptide, or directly fused to the adhesin.
  • DNA encoding an immunoglobulin light chain is typically coexpressed with the DNA encoding the adhesin-immunoglobulin heavy chain fusion protein.
  • the hybrid heavy chain and the light chain will be covalently associated to provide an immunoglobulin-like structure comprising two disulfide-linked immunoglobulin heavy chain-light chain pairs.
  • lmmunoadhesins are most conveniently constructed by fusing the cDNA sequence encoding the adhesin portion in-frame to an immunoglobulin cDNA sequence.
  • fusion to genomic immunoglobulin fragments can also be used (see, e.g. Aruffo et al, Cell 61:1303-1313 (1990); and Stamenkovic et al, Cell 66:1133-1144 (1991)).
  • the latter type of fusion requires the presence of Ig regulatory sequences for expression.
  • cDNAs encoding IgG heavy-chain constant regions can be isolated based on published sequences from cDNA libraries derived from spleen or peripheral blood lymphocytes, by hybridization or by polymerase chain reaction (PCR) techniques.
  • the cDNAs encoding the “adhesin” and the immunoglobulin parts of the immunoadhesin are inserted in tandem into a plasmid vector that directs efficient expression in the chosen host cells.
  • the invention also provides isolated nucleic acid encoding a polypeptide variant as disclosed herein, vectors and host cells comprising the nucleic acid, and recombinant techniques for the production of the polypeptide variant.
  • the nucleic acid encoding it is isolated and inserted into a replicable vector for further cloning (amplification of the DNA) or for expression.
  • DNA encoding the polypeptide variant is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the polypeptide variant).
  • Many vectors are available.
  • the vector components generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence.
  • the polypeptide variant of this invention may be produced recombinantly not only directly, but also as a fusion polypeptide with a heterologous polypeptide, which is preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide.
  • a heterologous polypeptide which is preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide.
  • the heterologous signal sequence selected preferably is one that is recognized and processed (ie., cleaved by a signal peptidase) by the host cell.
  • the signal sequence is substituted by a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II leaders.
  • a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II leaders.
  • yeast secretion the native signal sequence may be substituted by, e.g., the yeast invertase leader, ⁇ factor leader (including Saccharomyces and Kluyveromyces ⁇ -factor leaders), or acid phosphatase leader, the C. albicans glucoamylase leader, or the signal described in WO 90113646.
  • mammalian signal sequences as well as viral secretory leaders for example, the herpes simplex gD signal, are available.
  • the DNA for such precursor region is ligated in reading frame to DNA encoding the polypeptide variant.
  • Origin of Replication Component Both Expression and Cloning Vectors Contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells.
  • this sequence is one that enables the vector to replicate independently of the host chromosomal DNA, and includes origins of replication or autonomously replicating sequences.
  • origins of replication or autonomously replicating sequences are well known for a variety of bacteria, yeast, and viruses.
  • the origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria, the 2 ⁇ plasmid origin is suitable for yeast, and various viral origins (SV40, polyoma, adenovirus, VSV or BPV) are useful for cloning vectors in mammalian cells.
  • the origin of replication component is not needed for mammalian expression vectors (the SV40 origin may typically be used only because it contains the early promoter).
  • Expression and cloning vectors may contain a selection gene, also termed a selectable marker.
  • Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli.
  • One example of a selection scheme utilizes a drug to arrest growth of a host cell. Those cells that are successfully transformed with a heterologous gene produce a protein conferring drug resistance and thus survive the selection regimen. Examples of such dominant selection use the drugs neomycin, mycophenolic acid and hygromycin.
  • Suitable selectable markers for mammalian cells are those that enable the identification of cells competent to take up the polypeptide variant nucleic acid, such as DHFR, thymidine kinase, metallothionein-I and -II, preferably primate metallothionein genes, adenosine deaminase, ornithine decarboxylase, etc.
  • cells transformed with the DHFR selection gene are first identified by culturing all of the transformants in a culture medium that contains methotrexate (Mtx), a competitive antagonist of DHFR.
  • Mtx methotrexate
  • An appropriate host cell when wild-type DHFR is employed is the Chinese hamster ovary (CHO) cell line deficient in DHFR activity.
  • host cells transformed or co-transformed with DNA sequences encoding polypeptide variant, wild-type DHFR protein, and another selectable marker such as aminoglycoside 3′-phosphotransferase (APH) can be selected by cell growth in medium containing a selection agent for the selectable marker such as an aminoglycosidic antibiotic, e.g., kanamycin, neomycin, or G418. See U.S. Pat. No. 4,965,199.
  • APH aminoglycoside 3′-phosphotransferase
  • a suitable selection gene for use in yeast is the trpl gene present in the yeast plasmid YRp7 (Stinchcomb et al., Nature , 282:39 (1979)).
  • the trpl gene provides a selection marker for a mutant strain of yeast lacking the ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1. Jones, Genetics, 85:12 (1977).
  • the presence of the trp1 lesion in the yeast host cell genome then provides an effective environment for detecting transformation by growth in the absence of tryptophan.
  • Leu2-deficient yeast strains (ATCC 20,622 or 38,626) are complemented by known plasmids bearing the Leu2 gene.
  • vectors derived from the 1.6 ⁇ m circular plasmid pKDl can be used for transformation of Kluyveromyces yeasts.
  • an expression system for large-scale production of recombinant calf chymosin was reported for K. lactis. Van den Berg, Bio/Technology , 8:135 (1990).
  • Stable multi-copy expression vectors for secretion of mature recombinant human serum albumin by industrial strains of Kluyveromyces have also been disclosed. Fleer et al., Bio/Technology , 9:968-975 (1991).
  • Expression and cloning vectors usually contain a promoter that is recognized by the host organism and is operably linked to the polypeptide variant nucleic acid.
  • Promoters suitable for use with prokaryotic hosts include the phoA promoter, ⁇ -lactamase and lactose promoter systems, alkaline phosphatase, a tryptophan (trp) promoter system, and hybrid promoters such as the tac promoter.
  • trp tryptophan
  • Other known bacterial promoters are suitable. Promoters for use in bacterial systems also will contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA encoding the polypeptide variant.
  • S.D. Shine-Dalgarno
  • Promoter sequences are known for eukaryotes. Virtually all eukaryotic genes have an AT-rich region located approximately 25 to 30 bases upstream from the site where transcription is initiated. Another sequence found 70 to 80 bases upstream from the start of transcription of many genes is a CNCAAT region where N may be any nucleotide. At the 3′ end of most eukaryotic genes is an AATAAA sequence that may be the signal for addition of the poly A tail to the 3′ end of the coding sequence. All of these sequences are suitably inserted into eukaryotic expression vectors.
  • suitable promoting sequences for use with yeast hosts include the promoters for 3-phosphoglycerate kinase or other glycolytic enzymes, such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate kinase, triosephosphate isomerase, phosphoglucose isomerase, and glucokinase.
  • 3-phosphoglycerate kinase or other glycolytic enzymes such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate
  • yeast promoters which are inducible promoters having the additional advantage of transcription controlled by growth conditions, are the promoter regions for alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, degradative enzymes associated with nitrogen metabolism, metallothionein, glyceraldehyde-3-phosphate dehydrogenase, and enzymes responsible for maltose and galactose utilization.
  • Suitable vectors and promoters for use in yeast expression are further described in EP 73,657.
  • Yeast enhancers also are advantageously used with yeast promoters.
  • Polypeptide variant transcription from vectors in mammalian host cells is controlled, for example, by promoters obtained from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and most preferably Simian Virus 40 (SV40), from heterologous mammalian promoters, e.g., the actin promoter or an immunoglobulin promoter, from heat-shock promoters, provided such promoters are compatible with the host cell systems.
  • viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and most preferably Simian Virus 40 (
  • the early and late promoters of the SV40 virus are conveniently obtained as an SV40 restriction fragment that also contains the SV40 viral origin of replication.
  • the immediate early promoter of the human cytomegalovirus is conveniently obtained as a HindIII E restriction fragment.
  • a system for expressing DNA in mammalian hosts using the bovine papilloma virus as a vector is disclosed in U.S. Pat. No. 4,419,446. A modification of this system is described in U.S. Pat. No. 4,601,978.
  • Enhancer sequences are now known from mammalian genes (globin, elastase, albumin, ⁇ -fetoprotein, and insulin). Typically, however, one will use an enhancer from a eukaryotic cell virus. Examples include the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.
  • the enhancer may be spliced into the vector at a position 5′ or 3′ to the polypeptide variant-encoding sequence, but is preferably located at a site 5′ from the promoter.
  • Expression vectors used in eukaryotic host cells will also contain sequences necessary for the termination of transcription and for stabilizing the mRNA. Such sequences are commonly available from the 5′ and, occasionally 3′ , untranslated regions of eukaryotic or viral DNAs or cDNAs. These regions contain nucleotide segments transcribed as polyadenylated fragments in the untranslated portion of the mRNA encoding the polypeptide variant.
  • One useful transcription termination component is the bovine growth hormone polyadenylation region. See WO94/11026 and the expression vector disclosed therein.
  • Suitable host cells for cloning or expressing the DNA in the vectors herein are the prokaryote, yeast, or higher eukaryote cells described above.
  • Suitable prokaryotes for this purpose include eubacteria, such as Gram-negative or Gram-positive organisms, for example, Enterobacteriaceae such as Escherichia, e.g., E coli , Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium , Serratia, e.g., Serratia marcescans , and Shigella, as well as Bacilli such as B. subtilis and B.
  • Enterobacteriaceae such as Escherichia, e.g., E coli , Enterobacter, Erwinia, Klebsiella, Proteus
  • Salmonella e.g., Salmonella typhimurium
  • Serratia e.g
  • E. coil cloning host is E. coli 294 (ATCC 31,446), although other strains such as E. coli B, E. coil X1776 (ATCC 31,537), and E. coli W3110 (ATCC 27,325) are suitable. These examples are illustrative rather than limiting.
  • eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for polypeptide variant-encoding vectors.
  • Saccharomyces cerevisiae or common baker's yeast, is the most commonly used among lower eukaryotic host microorganisms.
  • a number of other genera, species, and strains are commonly available and useful herein, such as Schizosaccharomyces pombe; Kluyveromyces hosts such as, e.g., K lactis, K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K.
  • waltii ATCC 56,500
  • K. drosophilarum ATCC 36,906
  • K. thermotolerans K. marxianus
  • yarrowia EP 402,226
  • Pichia pastoris EP 183,070
  • Candida Trichoderma reesia
  • Neurospora crassa Schwanniomyces such as Schwanniomyces occidentalis
  • filamentous fungi such as, e.g., Neurospora, Penicillium, Tolypocladium, and Aspergillus hosts such as A. nidulans and A. niger .
  • Suitable host cells for the expression of glycosylated polypeptide variant are derived from multicellular organisms.
  • invertebrate cells include plant and insect cells.
  • Numerous baculoviral strains and variants and corresponding permissive insect host cells from hosts such as Spodoptera frugiperda (caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruiffly), and Bombyx mori have been identified.
  • a variety of viral strains for transfection are publicly available, e.g., the L-1 variant of Autographa californica NPV and the Bm-5 strain of Bombyx mori NPV, and such viruses may be used as the virus herein according to the present invention, particularly for transfection of Spodoptera frugiperda cells.
  • Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato, and tobacco can also be utilized as hosts.
  • vertebrate cells have been greatest in vertebrate cells, and propagation of vertebrate cells in culture (tissue culture) has become a routine procedure.
  • useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol . 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells/-DHFR (CHO, Urlaub et al, Proc. Natl. Acad. Sci. USA 77:4216 (1980)); mouse sertoli cells (TM4, Mather, Biol. Reprod .
  • COS-7 monkey kidney CV1 line transformed by SV40
  • human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol . 36:59 (1977)
  • monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N.Y. Acad. Sci . 383:44-68 (1982)); MRC 5 cells; FS4 cells; and a human hepatoma line (Hep G2).
  • Host cells are transformed with the above-described expression or cloning vectors for polypeptide variant production and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.
  • the host cells used to produce the polypeptide variant of this invention may be cultured in a variety of media.
  • Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPMI-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing the host cells.
  • any of these media may be supplemented as necessary with hormones and/or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics (such as GENTAMYCINTM drug), trace elements (defined as inorganic compounds usually present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements may also be included at appropriate concentrations that would be known to those skilled in the art.
  • the culture conditions such as temperature, pH, and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.
  • the polypeptide variant can be produced intracellularly, in the periplasmic space, or directly secreted into the medium. If the polypeptide variant is produced intracellularly, as a first step, the particulate debris, either host cells or lysed fragments, is removed, for example, by centrifugation or ultrafiltration. Carter et al., Bio/lTechnology 10:163-167 (1992) describe a procedure for isolating antibodies which are secreted to the periplasmic space of E coil .
  • cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30 min.
  • PMSF phenylmethylsulfonylfluoride
  • Cell debris can be removed by centrifugation.
  • supernatants from such expression systems are generally first concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit.
  • a protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants.
  • the polypeptide variant composition prepared from the cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being the preferred purification technique.
  • affinity chromatography is the preferred purification technique.
  • the suitability of protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc region that is present in the polypeptide variant. Protein A can be used to purify polypeptide variants that are based on human ⁇ 1, ⁇ 2, or ⁇ 4 heavy chains (Lindmark et al, J. Immunol. Meth . 62:1-13 (1983)).
  • Protein G is recommended for all mouse isotypes and for human ⁇ 3 (Guss et al, EMBO J . 5:15671575 (1986)).
  • the matrix to which the affinity ligand is attached is most often agarose, but other matrices are available. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose.
  • the polypeptide variant comprises a C H 3 domain
  • the Bakerbond ABXTM resin J. T. Baker, Phillipsburg, N.J.
  • the mixture comprising the polypeptide variant of interest and contaminants may be subjected to low pH hydrophobic interaction chromatography using an elution buffer at a pH between about 2.5-4.5, preferably performed at low salt concentrations (e.g.,from about 0-0.25M salt).
  • Therapeutic formulations of the polypeptide variant are prepared for storage by mixing the polypeptide variant having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers ( Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions.
  • Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyidimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arg
  • the formulation herein may also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other.
  • Such molecules are suitably present in combination in amounts that are effective for the purpose intended.
  • the active ingredients may also be entrapped in microcapsule prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsule and poly-(methylmethacylate) microcapsule, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions.
  • colloidal drug delivery systems for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules
  • formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.
  • Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the polypeptide variant, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No.
  • copolymers of L-glutamic acid and ⁇ ethyl-L-glutamate copolymers of L-glutamic acid and ⁇ ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOTTM (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-( ⁇ )-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods.
  • encapsulated antibodies When encapsulated antibodies remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37°C., resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S-S bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulihydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.
  • the polypeptide variant of the invention may be used as an affinity purification agent.
  • the polypeptide variant is immobilized on a solid phase such a Sephadex resin or filter paper, using methods well known in the art.
  • the immobilized polypeptide variant is contacted with a sample containing the antigen to be purified, and thereafter the support is washed with a suitable solvent that will remove substantially all the material in the sample except the antigen to be purified, which is bound to the immobilized polypeptide variant. Finally, the support is washed with another suitable solvent, such as glycine buffer, pH 5.0, that will release the antigen from the polypeptide variant.
  • polypeptide variant may also be useful in diagnostic assays, e.g., for detecting expression of an antigen of interest in specific cells, tissues, or serum.
  • polypeptide variant typically will be labeled with a detectable moiety.
  • labels are available which can be generally grouped into the following categories:
  • Radioisotopes such as 35 S, 14 C, 125 I, 3 H, and 131 I.
  • the polypeptide variant can be labeled with the radioisotope using the techniques described in Current Protocots in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley-lnterscience, New York, New York, Pubs. (1991) for example and radioactivity can be measured using scintillation counting.
  • Fluorescent labels such as rare earth chelates (europium chelates) or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin and Texas Red are available.
  • the fluorescent labels can be conjugated to the polypeptide variant using the techniques disclosed in Current Protocols in Immunology, supra, for example. Fluorescence can be quantified using a fluorimeter.
  • the enzyme generally catalyzes a chemical alteration of the chromogenic substrate that can be measured using various techniques. For example, the enzyme may catalyze a color change in a substrate, which can be measured spectrophotometrically. Alternatively, the enzyme may alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying a change in fluorescence are described above.
  • the chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light which can be measured (using a chemiluminometer, for example) or donates energy to a fluorescent acceptor.
  • enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase; U.S. Pat. No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, ⁇ -galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like.
  • luciferases e.g., firefly luciferase and bacterial lucifera
  • enzyme-substrate combinations include, for example:
  • HRPO Horseradish peroxidase
  • OPD orthophenylene diamine
  • TMB 3,3′,5,5′-tetramethyl benzidine hydrochloride
  • ⁇ -D-galactosidase ( ⁇ -D-Gal) with a chromogenic substrate (e.g., p-nitrophenyl- ⁇ -D-galactosidase) or fluorogenic substrate 4-methylumbelliferyl- ⁇ -D-galactosidase.
  • a chromogenic substrate e.g., p-nitrophenyl- ⁇ -D-galactosidase
  • fluorogenic substrate 4-methylumbelliferyl- ⁇ -D-galactosidase
  • the label is indirectly conjugated with the polypeptide variant.
  • the polypeptide variant can be conjugated with biotin and any of the three broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the polypeptide variant in this indirect manner.
  • the polypeptide variant is conjugated with a small hapten (e.g., digoxin) and one of the different types of labels mentioned above is conjugated with an anti-hapten polypeptide variant (e.g., anti-digoxin antibody).
  • an anti-hapten polypeptide variant e.g., anti-digoxin antibody
  • the polypeptide variant need not be labeled, and the presence thereof can be detected using a labeled antibody which binds to the polypeptide variant.
  • polypeptide variant of the present invention may be employed in any known assay method, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, Monoclonal Antibodies:A Manualof Techniques , pp.147-158 (CRC Press, Inc. 1987).
  • the polypeptide variant may also be used for in vivo diagnostic assays.
  • the polypeptide variant is labeled with a radionuclide (such as 111 In, 99 Tc, 14 C, 131 I, 125 I, 3 H, 32 P or 35 S) so that the antigen or cells expressing it can be localized using immunoscintiography.
  • a radionuclide such as 111 In, 99 Tc, 14 C, 131 I, 125 I, 3 H, 32 P or 35 S
  • the polypeptide variant of the present invention may be used to treat a mammal e.g. a patient suffering from a disease or disorder who could benefit from administration of the polypeptide variant.
  • the conditions which can be treated with the polypeptide variant are many and include cancer (e.g. where the polypeptide variant binds the HER2 receptor or CD20); allergic conditions such as asthma (with an anti-IgE antibody); and LFA-mediated disorders (e.g. where the polypeptide variant is an anti-LFA-1 or anti-ICAM-1 antibody) etc.
  • exemplary diseases or disorders to be treated include: cancer (e.g.
  • ADCC function is desirable, but complement activation would lead to amplified side effects, such as vasculitis in the blood vessels at the tumor site); disorders treated with an agonist antibody; disorders wherein the polypeptide variant binds a soluble antigen and wherein stoichiometry leads to immune complexes which activate the complement cascade and result in unwanted side effects; conditions employing an antagonist antibody which downmodulates receptor function without damaging tissue or organ function;
  • the polypeptide variant is administered by any suitable means, including parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal, and, if desired for local immunosuppressive treatment, intralesional administration.
  • Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
  • the polypeptide variant is suitably administered by pulse infusion, particularly with declining doses of the polypeptide variant.
  • the dosing is given by injections, most preferably intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic.
  • polypeptide variant For the prevention or treatment of disease, the appropriate dosage of polypeptide variant will depend on the type of disease to be treated, the severity and course of the disease, whether the polypeptide variant is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the polypeptide variant, and the discretion of the attending physician.
  • the polypeptide variant is suitably administered to the patient at one time or over a series of treatments.
  • polypeptide variant is an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion.
  • a typical daily dosage might range from about 1 ⁇ g/kg to 100 mg/kg or more, depending on the factors mentioned above.
  • the treatment is sustained until a desired suppression of disease symptoms occurs.
  • other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays.
  • the polypeptide variant composition will be formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners.
  • the “therapeutically effective amount” of the polypeptide variant to be administered will be governed by such considerations, and is the minimum amount necessary to prevent, ameliorate, or treat a disease or disorder.
  • the polypeptide variant need not be, but is optionally formulated with one or more agents currently used to prevent or treat the disorder in question.
  • the effective amount of such other agents depends on the amount of polypeptide variant present in the formulation, the type of disorder or treatment, and other factors discussed above. These are generally used in the same dosages and with administration routes as used hereinbefore or about from 1 to 99% of the heretofore employed dosages.
  • This assay determines binding of an IgG Fc region to recombinant Fc ⁇ RIIa, Fc ⁇ RIIb and Fc ⁇ RIIIa ⁇ subunits expressed as His6-glutathione S transferase (GST)-tagged fusion proteins. Since the affinity of the Fc region of IgG1 for the Fc ⁇ RI is in the nanomolar range, the binding of IgG1 Fc mutants can be measured by titrating monomeric IgG and measuring bound IgG with a polyclonal anti-lgG in a standard ELISA format (Example 2 below). The affinity of the other members of the Fc ⁇ R family, ie. Fc ⁇ RIIa, Fc ⁇ RIIb and Fc ⁇ RIIIa for IgG is however in the micromolar range and binding of monomeric IgG1 for these receptors can not be reliably measured in an ELISA format.
  • GST His6-glutathione S transferase
  • the following assay utilizes Fc mutants of recombinant anti-IgE E27 (FIGS. 4A and 4B) which, when mixed with human IgE at a 1:1 molar ratio, forms a stable hexamer consisting of three anti-IgE molecules and three IgE molecules.
  • a recombinant chimeric form of IgE (chimeric IgE) was engineered and consists of a human IgE Fc region and the Fab of an anti-VEGF antibody (Presta et al Cancer Research 57:4593-4599 (1997)) which binds two VEGF molecules per mole of anti-VEGF.
  • VEGF vascular endothelial growth factor
  • hexamers When recombinant human VEGF is added at a 2:1 molar ratio to chimeric IgE:E27 hexamers, the hexamers are linked into larger molecular weight complexes via the chimeric IgE Fab:VEGF interaction.
  • the E27 component of this complex binds to the Fc ⁇ RIIa, Fc ⁇ RIIb and Fc ⁇ IIIa ⁇ subunits with higher avidity to permit detection in an ELISA format.
  • Receptor Coat. Fc ⁇ receptor ⁇ subunits were expressed as GST fusions of His6 tagged extracellular domains (ECDs) in 293 cells resulting in an ECD-6His-GST fusion protein (Graham et al J. Gen. Virol . 36:59-72 (1977) and Gorman et al DNA Prot. Eng. Tech . 2:3-10 (1990)) and purified by Ni-NTA column chromatography (Qiagen, Australia) and buffer exchanged into phosphate buffered saline (PBS). Concentrations were determined by absorption at 280nm using extinction coefficients derived by amino acid composition analysis. Receptors were coated onto Nunc F96 maxisorb plates (cat no.
  • E27:chimeric IgE:VEGF(1:1:2 molar ratio) complexes are added to Fc ⁇ R ⁇ subunit coated plates at E27 concentrations of 5 ⁇ g and 1 ⁇ g total IgG in quadruplicate in assay buffer and incubated for 120 minutes at 25° C. on an orbital shaker.
  • HRP horse radish peroxidase conjugated goat anti-human IgG ( ⁇ ) heavy chain specific
  • Plates are washed 5 ⁇ with wash buffer to remove unbound HRP goat anti-human IgG and bound anti-lgG is detected by adding 100 ⁇ l of substrate solution (0.4mg/ml o-phenylenedaimine dihydrochloride, Sigma P6912, 6 mM H 2 O 2 in PBS) and incubating for 8 min at 25° C. Enzymatic reaction is stopped by the addition of 100 ⁇ l 4.5N H 2 SO 4 and colorimetric product is measured at 490 nm on a 96 well plate densitometer (Molecular Devices). Binding of E27 mutant complexes is expressed as a percent of the wild type E27 containing complex.
  • the heavy and light chain plasmids were co-transfected into an adenovirus transformed human embryonic kidney cell line as previously described (Werther et al., J. Immunol 157:4986 (1996)). The media was changed to serum-free 24 hours after transfection and the secreted antibody was harvested after five days.
  • the antibodies were purified using Protein A-SEPHAROSE CL-4BTM (Pharmacia), buffer exchanged and concentrated to 0.5 ml with PBS using a Centricon-30 (Amicon), and stored at 4° C. The concentration of the antibody was determined using total Ig-binding ELISA.
  • C1q Binding ELISA Costar 96 well plates were coated overnight at 4° C. with the indicated concentrations of 2B8 in coating buffer (0.05 M sodium carbonate buffer), pH 9. The plates were then washed 3 ⁇ with PBS/ 0.05% TWEEN 20TM, pH 7.4 and blocked with 200 ⁇ l of ELISA diluent without thimerosal (0.1 M NaPO4/0.1M NaCl/0.1% gelatin/0.05% TWEEN 20 TM/ 0.05% ProClin300) for 1 hr at room temperature.
  • the plate was washed 3 ⁇ with wash buffer, an aliquot of 100 ⁇ l of 2 ⁇ g/ml C1q (Quidel, San Diego, Calif.) was added to each well and incubated for 2 hrs at room temperature. The plate was then washed 6 ⁇ with wash buffer. 100 ⁇ l of a 1:1000 dilution of sheep anti-complement C1q peroxidase conjugated antibody (Biodesign) was added to each well and incubated for 1 hour at room temperature. The plate was again washed 6 ⁇ with wash buffer and 100 ⁇ l of substrate buffer (PBS/0.012% H 2 O 2 ) containing OPD (O-phenylenediamine dihydrochloride (Sigma)) was added to each well.
  • substrate buffer PBS/0.012% H 2 O 2
  • OPD O-phenylenediamine dihydrochloride
  • the oxidation reaction observed by the appearance of a yellow color, was allowed to proceed for 30 minutes and stopped by the addition of 100 ⁇ l of 4.5 N H 2 SO 4 .
  • the absorbance was then read at (492-405) nm using a microplate reader (SPECTRA MAX 250TM, Molecular Devices Corp.).
  • the appropriate controls were run in parallel (i.e. the ELISA was performed without C1q for each concentration of C2B8 used and also the ELISA was performed without C2B8).
  • C1q binding was measured by plotting the absorbance (492-405) nm versus concentration of C2B8 in ⁇ g/ml using a 4-parameter curve fitting program (KALEIDAGRAPHTM) and comparing EC 50 values.
  • Human complement (Quidel) was diluted 1:3 in RHB buffer and WIL2-S cells (available from the ATCC, Manassas, Va.) which express the CD20 antigen were diluted to a density of 1 ⁇ 10 6 cells /ml with RHB buffer.
  • Mixtures of 150 ⁇ l containing equal volumes of C2B8, diluted human complement and WIL2-S cells were added to a flat bottom tissue culture 96 well plate and allowed to incubate for 2 hrs at 37° C. and 5% CO 2 to facilitate complement mediated cell lysis. 50 ⁇ l of alamar blue (Accumed International) was then added to each well and incubated overnight at 37° C.
  • the absorbance was measured using a 96-well fluorometer with excitation at 530 nm and emission at 590 nm. As described by Gazzano-Santoro et al., the results are expressed in relative fluorescence units (RFU). The sample concentrations were computed from a C2B8 standard curve and the percent activity as compared to wild type C2B8 is reported for each mutant.
  • CD20 Binding Potency of the C2B8 Mutants The binding of C2B8 and mutants to the CD20 antigen were assessed by a method previously described (Reff et al, (1994), supra; reviewed in Gazzano-Santoro et al, (1996), supra). WIL2-S cells were grown for 3-4 days to a cell density of 1 ⁇ 10 6 cells/ml. The cells were washed and spun twice in FACS buffer (PBS/0.1% BSA/0.02% NaN 3 ) and resuspended to a cell density of 5 ⁇ 10 6 cells/ml.
  • FACS buffer PBS/0.1% BSA/0.02% NaN 3
  • Fc ⁇ RI ⁇ subunit-GST fusion was coated onto Nunc F96 maxisorb plates (cat no. 439454) by adding 100 ⁇ l of receptor-GST fusion at 1 ⁇ g/ml in PBS and incubated for 48 hours at 4° C. Prior to assay, plates are washed 3 ⁇ with 250 ⁇ l of wash buffer (PBS pH 7.4 containing 0.5% TWEEN 20TM) and blocked with 250 ⁇ l of assay buffer (50 mM Tris buffered saline, 0.05% TWEEN 20TM, 0.5% RIA grade bovine albumin (Sigma A7888), and 2 mM EDTA pH 7.4).
  • wash buffer PBS pH 7.4 containing 0.5% TWEEN 20TM
  • assay buffer 50 mM Tris buffered saline, 0.05% TWEEN 20TM, 0.5% RIA grade bovine albumin (Sigma A7888), and 2 mM EDTA pH 7.4
  • Plates are washed 5 ⁇ with wash buffer to remove unbound HRP goat anti-human IgG and bound anti-lgG is detected by adding 100 ⁇ l of substrate solution (0.4mg/ml o-phenylenedaimine dihydrochloride, Sigma P6912, 6 mM H 2 O 2 in PBS) and incubating for 8 min at 25° C. Enzymatic reaction is stopped by the addition of 100 ⁇ l 4.5N H 2 SO 4 and colorimetric product is measured at 490 nm on a 96 well plate densitometer (Molecular Devices). Binding of variant is expressed as a percent of the wild type molecule.
  • Fc ⁇ RII and III binding ELISAs were performed as described in Example 1 above.
  • the IgG2 mutant appears to have a much lower affinity for C1q than the E318A and K320A mutants (FIG. 2).
  • the results demonstrate that E318 and K320 do not constitute the core C1q binding sites for human IgG1.
  • the K322A substitution had a significant effect on both complement activity and C1q binding.
  • the K322A mutant did not have CDC activity when tested in the above CDC assay and was more than a 100 fold lower than wild type C2B8 in binding to C1q (FIG. 2).
  • K322 is the only residue of the proposed core C1q binding sites that appeared to have a significant effect on complement activation and C1q binding.
  • the mutation P329A results in an antibody that does not activate complement and is more than a 100 fold lower in binding to C1q (FIG. 3) when compared to wild type C2B8 (FIG. 2).

Abstract

A variant of a polypeptide comprising a human IgG Fc region is described, which variant comprises an amino acid substitution at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human IgG Fc region. Such variants display altered effector function. For example, Clq binding and/or complement dependent cytotoxicity (CDC) activity may be reduced or abolished in the variant polypeptide. The application also describes an immune complex and an assay for determining binding of an analyte, such as an Fc region-containing polypeptide, to a receptor.

Description

  • This is a non-provisional application claiming priority under 35 USC §119 to provisional application No. 60/080,447 filed Apr. 2, 1998, the entire disclosure of which is hereby incorporated by reference.[0001]
    • BACKGROUND OF THE INVENTION
  • 1. Field of the Invention [0002]
  • The present invention concerns variants of polypeptides comprising an Fc region. More particularly, the present invention concerns Fc region-containing polypeptides that have altered effector function as a consequence of one or more amino acid substitutions in the Fc region of the nonvariant polypeptide. The invention also relates to novel immune complexes and an assay for determining binding of an analyte, such as an Fc region-containing polypeptide, to a receptor. [0003]
  • 2. Description of Related Art [0004]
  • Antibodies are proteins, which exhibit binding specificity to a specific antigen. Native antibodies are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies between the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (V[0005] H) followed by a number of constant domains. Each light chain has a variable domain at one end (VL) and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light and heavy chain variable domains.
  • The term “variable” refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies and are responsible for the binding specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed through the variable domains of antibodies. It is concentrated in three segments called complementarity determining regions (CDRs) both in the light chain and the heavy chain variable domains. The more highly conserved portions of the variable domains are called the framework regions (FR). The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a βsheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the β-sheet structure. The CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen binding site of antibodies (see Kabat et al., Sequences of [0006] Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).
  • The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions. Depending on the amino acid sequence of the constant region of their heavy chains, antibodies or immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, and several of these may be further divided into subclasses (isotypes), e.g. IgG1, IgG2, IgG3, and IgG4; IgA1 and IgA2. The heavy chain constant regions that correspond to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. Of the various human immunoglobulin classes, only human IgG1, IgG2, IgG3 and IgM are known to activate complement. [0007]
  • A schematic representation of the native IgG1 structure is shown in FIG. 1, where the various portions of the native antibody molecule are indicated. Papain digestion of antibodies produces two identical antigen binding fragments, called Fab fragments, each with a single antigen binding site, and a residual “Fc” fragment, whose name reflects its ability to crystallize readily. The crystal structure of the human IgG Fc region has been determined (Deisenhofer, [0008] Biochemistry 20:2361-2370 (1981)). In human IgG molecules the Fc region is generated by papain cleavage N-terminal to Cys 226. The Fc region is central to the effector functions of antibodies.
  • The effector functions mediated by the antibody Fc region can be divided into two categories: (1) effector functions that operate after the binding of antibody to an antigen (these functions involve the participation of the complement cascade or Fc receptor (FcR)-bearing cells); and (2) effector functions that operate independently of antigen binding (these functions confer persistence in the circulation and the ability to be transferred across cellular barriers by transcytosis). Ward and Ghetie, [0009] Therapeutic Immunology 2:77-94 (1995).
  • While binding of an antibody to the requisite antigen has a neutralizing effect that might prevent the binding of a foreign antigen to its endogenous target (e.g. receptor or ligand), binding alone may not remove the foreign antigen. To be efficient in removing and/or destructing foreign antigens, an antibody should be endowed with both high affinity binding to its antigen, and efficient effector functions. [0010]
    • C1q Binding
  • C1q and two serine proteases, C1r and C1s, form the complex C1, the first component of the complement dependent cytotoxicity (CDC) pathway. C1q is a hexavalent molecule with a molecular weight of approximately 460,000 and a structure likened to a bouquet of tulips in which six collagenous “stalks” are connected to six globular head regions. Burton and Woof, [0011] Advances in Immunol. 51:1-84 (1992). To activate the complement cascade, it is necessary for C1q to bind to at least two molecules of IgG1, IgG2, or IgG3 (the consensus is that IgG4 does not activate complement), but only one molecule of IgM, attached to the antigenic target. Ward and Ghetie, Therapeutic Immunology 2:77-94 (1995) at page 80.
  • Based upon the results of chemical modifications and crystallographic studies, Burton et aL ([0012] Nature, 288:338-344 (1980)) proposed that the binding site for the complement subcomponent C1q on IgG involves the last two (C-terminal) β-strands of the CH2 domain. Burton later suggested (Molec. Immunol., 22(3):161-206 (1985)) that the region comprising amino acid residues 318 to 337 might be involved in complement fixation.
  • Duncan and Winter ([0013] Nature 332:738-40 (1988)), using site directed mutagenesis, reported that Glu318, Lys320 and Lys322 form the binding site to C1q. The data of Duncan and Winter were generated by testing the binding of a mouse IgG2b isotype to guinea pig C1q. The role of Glu318, Lys320 and Lys322 residues in the binding of C1q was confirmed by the ability of a short synthetic peptide containing these residues to inhibit complement mediated lysis. Similar results are disclosed in U.S. Pat. No. 5,648,260 issued on Jul. 15, 1997, and U.S. Pat. No. 5,624,821 issued on Apr. 29, 1997.
  • The residue Pro331 has been implicated in C1q binding by analysis of the ability of human IgG subclasses to carry out complement mediated cell lysis. Mutation of Ser331 to Pro331 in IgG4 conferred the ability to activate complement. (Tao et aL, J. [0014] Exp. Med., 178:661-667 (1993); Brekke et al., Eur. J. lmmunol, 24:2542-47 (1994)).
  • From the comparison of the data of the Winter group, and the Tao et al. and Brekke et al. papers, Ward and Ghetie concluded in their review article that there are at least two different regions involved in the binding of C1q: one on the β-strand of the CH2 domain bearing the Glu318, Lys320 and Lys322 residues, and the other on a turn located in close proximity to the sameβ-strand, and containing a key amino acid residue at position 331. [0015]
  • Other reports suggested that human IgG1 residues Lys235, and Gly237, located in the lower hinge region, play a critical role in complement fixation and activation. Xu et al., [0016] J. Immunol. 150:152A (Abstract) (1993). WO94/29351 published Dec. 22, 1994 reports that amino acid residues necessary for C1q and FcR binding of human IgG1 are located in the N-terminal region of the CH2 domain, i.e. residues 231 to 238.
  • It has further been proposed that the ability of IgG to bind C1q and activate the complement cascade also depends on the presence, absence or modification of the carbohydrate moiety positioned between the two CH2 domains (which is normally anchored at Asn297). Ward and Ghetie, [0017] Therapeutic Immunology 2:77-94 (1995) at page 81.
    • Fc Receptor Binding
  • Effector functions can also be mediated by the interaction of the Fc region of an antibody with Fc receptors (FcRs), which are specialized cell surface receptors on hematopoietic cells. Fc receptors belong in the immunoglobulin superfamily, and have been shown to mediate both the removal of antibody-coated pathogens by phagocytosis of immune complexes, and the lysing of erythrocytes and various other cellular targets (e.g. tumor cells) coated with the corresponding antibody, via antibody dependent cell mediated cytotoxicity (ADCC). Van de Winkel and Anderson, [0018] J. Leuk. Biol. 49:511-24 (1991).
  • FcRs are defined by their specificity for immunoglobulin isotypes; Fc receptors for IgG antibodies are referred to as FcγR, for IgE as FcεR, for IgA as FcαR and so on. Three subclasses of gamma receptors have been identified: FcεRI (CD64), FcγRII (CD32) and FcγRIII (CD16). Because each FcγR subclass is encoded by two or three genes, and alternative RNA spicing leads to multiple transcripts, a broad diversity in FcγR isoforms exists. The three genes encoding the FcγRI subclass (FcγRIA, FcγRIB and FcγRIC) are clustered in region 1q21.1 of the long arm of [0019] chromosome 1; the genes encoding FcγRII isoforms (FcγRIIA, FcγRIIB and FcγRIIC) and the two genes encoding FcγRIII (FcγRIIIA and FcγRIIIB) are all clustered in region 1q22. FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et aL, Immunomethods 4:25-34 (1994); and de Haas etal., J. Lab. Clin. Med. 126:330-41 (1995).
  • While FcγRI binds monomeric IgG with a high affinity, FcγRII and FcγRIII are low-affinity receptors, interacting with complexed or aggregated IgG. The classical method for detecting these low-affinity receptors is by “rosetting” using antibody-coated erythrocytes (EA) sensitized with IgGs. Bredius et aL evaluated rosette formation between IgG-sensitized red blood cells and polymorphonuclear leukocytes (PMN) which express FcγRIIa and FcγRIIIb at their cell-surface. Rosette was defined as three of more EA bound per PMN (Bredius etal. [0020] Immunology 83:624-630 (1994)). See, also, Tax et aL J. Immunol. 133(3):1185-1189 (1984); Nagarajan et aL J. Biol. Chem. 270(43):25762-25770 (1995); and Warmerdam et al. J. Immunol. 147(4):1338-1343 (1991concerning rosette assays. However, binding of these EA “immune complexes” to FcR is not easily quantified. Accordingly, more defined complexes with detectable affinity for these FcRs have been developed. For example, IgG dimers have been formed using anti-light chain monoclonal antibodies (Nagarajan et al, supra and Warmerdam et al., supra) or chemical cross-linking agents (Hogarth et al. Immunomethods 4:17-24 (1994); and Tamm et aL J. Biol Chem. 271(7):3659-3666(1996)). Heat-aggregated immune complexes have also been evaluated for binding to cells expressing FcRs (Tax et al., supra and Tam et al., supra).
  • The binding site for the FcγRs on human IgG was found to reside in the lower hinge region, primarily involving residues at amino acid positions 233-238, all of which were found to be necessary for full FcγR binding activity. Canfield and Morrison, [0021] J. Exp. Med. 173:1483-91 (1991); Chappel et al., Proc. Natl. Acad. Sci. USA, 88:9036-40 (1991); Lund etal., J. Immunol., 147:2657-62 (1991); Lund et al, Molec. ImmunoL., 29:53-59 (1992); Jefferis et al., Molec. Immunol., 27:1237-40 (1990); and Sarmay et al., Molec. Immunol, 29:633-639 (1992).
  • Pro331 in IgG3 was changed to Ser, and the affinity of this mutant to target cells analyzed. The affinity was found to be six-fold lower than that of unmutated IgG3, indicating the involvement of Pro331 in FcγRI binding. Morrison et al., [0022] Immunologist, 2:119-124 (1994); and Canfield and Morrison, J. Exp. Med. 173:1483-91 (1991).
  • In addition, Glu318 was identified as being involved in binding to FcγRII. Ward and Ghetie, [0023] Therapeutic Immunology 2:77-94 (1995).
    • SUMMARY OF THE INVENTION
  • The present invention provides a variant of a polypeptide comprising a human IgG Fc region, which variant comprises an amino acid substitution at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat. [0024]
  • The invention further relates to a variant of a polypeptide comprising a human IgG Fc region, which variant binds FcγRI, FcγRII, FcγRIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 322 or amino acid position 329, or both amino acid positions of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat. [0025]
  • In yet a further aspect, the invention provides a method for modifying a polypeptide comprising a human IgG Fc region comprising substituting an amino acid residue at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human lgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat. [0026]
  • The invention also provides a composition comprising the polypeptide variant and a physiologically acceptable carrier or diluent. This composition for potential therapeutic use is sterile and may be lyophilized. [0027]
  • Diagnostic and therapeutic uses for the polypeptide variant are contemplated. In one diagnostic application, the invention provides a method for determining the presence of a protein of interest comprising exposing a sample suspected of containing the protein to the polypeptide variant and determining binding of the polypeptide variant to the sample. In one therapeutic application, the invention provides a method of treating a mammal suffering from a disorder comprising administering to the mammal a therapeutically effective amount of a variant of a polypeptide comprising a human IgG Fc region, which variant binds FcγRI, FcγRII, FcγRIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 329, 331 or 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat. [0028]
  • The invention further provides: isolated nucleic acid encoding the polypeptide variant; a vector comprising the nucleic acid, optionally, operably linked to control sequences recognized by a host cell transformed with the vector; a host cell comprising the vector; a process for producing the polypeptide variant comprising culturing this host cell so that the nucleic acid is expressed and, optionally, recovering the polypeptide variant from the host cell culture (e.g. from the host cell culture medium). [0029]
  • The invention also pertains to an immune complex comprising: (a) an Fc region-containing polypeptide; (b) a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide; and (c) a second target molecule comprises at least two binding sites for the first target molecule. The immune complex may be used in an FcR-binding assay, particularly where the FcR has a low affinity for the Fc region-containing polypeptide. Other uses for the immune complex are disclosed herein. [0030]
  • Moreover, the invention provides a method for determining binding of an analyte, such as an Fc region-containing polypeptide, to a receptor (e.g. a low affinity FcR) comprising the following steps performed sequentially: (a) forming a molecular complex between the analyte and a first target molecule, wherein the first target molecule comprises at least two binding sites for the analyte; and (b) determining binding of the molecular complex of step (a) to the receptor (e.g. to a binding domain of the receptor coated on an assay plate). Optionally, the molecular complex of step (a) further comprises a second target molecule which comprises at least two binding sites for the first target molecule. [0031]
  • The invention also relates to an assay kit, such as a kit useful for determining binding of an analyte to a receptor comprising: (a) a first target molecule which comprises at least two binding sites for the analyte; and (b) a second target molecule which comprises at least two binding sites for the first target molecule.[0032]
    • BRIEF DESCRIPTIOM OF THE DRAWINDS
  • FIG. 1 is a schematic representation of a native IgG. Disulfide bonds are represented by heavy lines between CH1 and CL domains and the two CH2 domains. V is variable domain; C is constant domain; L stands for light chain and H stands for heavy chain. [0033]
  • FIG. 2 shows C1q binding of wild type (wt) C2B8 antibody; C2B8 antibody with a human IgG2 constant region (IgG2); and mutants K322A, K320A and E318A. [0034]
  • FIG. 3 depicts C1q binding of mutants P331A, P329A and K322A. [0035]
  • FIGS. 4A (SEQ ID NO:1) and 4B (SEQ ID NO:2) depict the amino acid sequences of E27 anti-IgE antibody light chain (FIG. 4A) and heavy chain (FIG. 4B). [0036]
  • FIG. 5 is a schematic diagram of the “immune complex” prepared for use in the FcR assay described in Example 1. The hexamer comprising three anti-IgE antibody molecules (the “Fc region-containing polypeptide”) and three IgE molecules (the “first target molecule”) is shown. IgE has two “binding sites” for the anti-lgE antibody (E27) in the Fc region thereof. Each IgE molecule in the complex is further able to bind two VEGF molecules (“the second target polypeptide”). VEGF has two “binding sites” for IgE.[0037]
    • DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
  • I. Definitions [0038]
  • Throughout the present specification and claims, the numbering of the residues in an IgG heavy chain is that of the EU index as in Kabat et al., [0039] Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991), expressly incorporated herein by reference. The “EU index as in Kabat” refers to the residue numbering of the human IgG1 EU antibody.
  • The term “Fc region” is used to define a C-terminal region of an IgG heavy chain as shown FIG. 1. Although the boundaries of the Fc region of an IgG heavy chain might vary slightly, the human IgG heavy chain Fc region is usually defined to stretch from amino acid residue at position Cys226 to the carboxyl-terminus. The term “Fc region-containing polypeptide” refers to a polypeptide, such as an antibody or immunoadhesin (see definitions below), which comprises an Fc region. [0040]
  • The Fc region of an IgG comprises two constant domains, CH2 and CH3, as shown in FIG. 1. The “CH2” domain of a human IgG Fc region (also referred to as “Cγ2” domain) usually extends from amino acid 231 to amino acid 340. The CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgG molecule. It has been speculated that the carbohydrate may provide a substitute for the domain-domain pairing and help stabilize the CH2 domain. Burton, [0041] Molec. Immunol. 22:161-206 (1985).
  • “Hinge region” is generally defined as stretching from Glu216 to Pro230 of human IgG1 (Burton, [0042] Molec. Immunol. 22:161-206 (1985)) Hinge regions of other IgG isotypes may be aligned with the IgG1 sequence by placing the first and last cysteine residues forming inter-heavy chain S-S bonds in the same positions.
  • “C1q” is a polypeptide that includes a binding site for the Fc region of an immunoglobulin. C1q together with two serine proteases, C1r and C1s, forms the complex C1, the first component of the complement dependent cytotoxicity (CDC) pathway. Human C1q can be purchased commercially from, e.g. Quidel, San Diego, CA. [0043]
  • The term “Fc receptor” or “FcR” is used to describe a receptor that binds to the Fc region of an antibody. The preferred FcR is one, which binds an IgG antibody (a gamma receptor) and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced forms of these receptors. FcRs are reviewed in Ravetch and Kinet, [0044] Annu. Rev. Immunol 9:457-92 (1991); Capel et al, Immunomethods 4:25-34 (1994); and de Haas et al, J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term “FcR” herein. The term also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)).
  • The term “binding domain” refers to the region of a polypeptide that binds to another molecule. In the case of an FcR, the binding domain can comprise a portion of a polypeptide chain thereof (e.g. the α chain thereof) which is responsible for binding an Fc region. One useful binding domain is the extracellular domain of an FcR α chain. [0045]
  • The term “antibody” is used in the broadest sense and specifically covers monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments so long as they exhibit the desired biological activity. [0046]
  • “Antibody fragments”, as defined for the purpose of the present invention, comprise a portion of an intact antibody, generally including the antigen binding or variable region of the intact antibody, so long as they retain at least the CH2 region of an IgG immunoglobulin heavy chain constant domain, comprising amino acid residues 322, 329 and 331, and have the ability, alone or in combination with another antibody fragment, to specifically bind a selected antigen. Examples of antibody fragments include linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. The antibody fragments preferably retain at least part of the hinge and optionally the CH1 region of an IgG heavy chain. More preferably, the antibody fragments retain the entire constant region of an IgG heavy chain, and include an IgG light chain. [0047]
  • The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional (polyclonal) antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al, [0048] Nature 256:495 (1975), or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature 352:624-628 (1991) and Marks et al., J. Mol. Biol. 222:581-597 (1991), for example.
  • The monoclonal antibodies herein specifically include “chimeric” antibodies (immunoglobulins) in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al, [0049] Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)).
  • “Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity. In some instances, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., [0050] Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).
  • The term “hypervariable region” when used herein refers to the amino acid residues of an antibody which are responsible for antigen-binding. The hypervariable region comprises amino acid residues from a “complementarity determining region” or “CDR” (i.e. residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2) and 95-102 (H3) in the heavy chain variable domain; Kabat et al., [0051] Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)) and/or those residues from a “hypervariable loop” (i.e. residues 26-32 (L1), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (H1), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain; Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). “Framework” or “FR” residues are those variable domain residues other than the hypervariable region residues as herein defined.
  • As used herein, the term “immunoadhesin” designates antibody-like molecules which combine the “binding domains” of a heterologous “adhesin” protein (e.g. a receptor, ligand or enzyme) with an immunoglobulin constant domain. Structurally, the immunoadhesins comprise a fusion of the adhesin amino acid sequence with the desired binding specificity which is other than the antigen recognition and binding site (antigen combining site) of an antibody (i.e. is “heterologous”) and an immunoglobulin constant domain sequence. [0052]
  • The term “ligand binding domain” as used herein refers to any native cell-surface receptor or any region or derivative thereof retaining at least a qualitative ligand binding ability of a corresponding native receptor. In a specific embodiment, the receptor is from a cell-surface polypeptide having an extracellular domain that is homologous to a member of the immunoglobulin supergenefamily. Other receptors, which are not members of the immunoglobulin supergenefamily but are nonetheless specifically covered by this definition, are receptors for cytokines, and in particular receptors with tyrosine kinase activity (receptor tyrosine kinases), members of the hematopoietin and nerve growth factor receptor superfamilies, and cell adhesion molecules, e.g. (E-, L- and P-) selectins. [0053]
  • The term “receptor binding domain” is used to designate any native ligand for a receptor, including cell adhesion molecules, or any region or derivative of such native ligand retaining at least a qualitative receptor binding ability of a corresponding native ligand. This definition, among others, specifically includes binding sequences from ligands for the above-mentioned receptors. [0054]
  • An “antibody-immunoadhesin chimera” comprises a molecule that combines at least one binding domain of an antibody (as herein defined) with at least one immunoadhesin (as defined in this application). Exemplary antibody-immunoadhesin chimeras are the bispecific CD4-lgG chimeras described in Berg et al, [0055] PNAS (USA) 88:4723-4727 (1991) and Chamow et al, J. Immunol. 153:4268 (1994).
  • An “isolated” polypeptide is one that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the polypeptide, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. In preferred embodiments, the polypeptide will be purified (1) to greater than 95% by weight of polypeptide as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain. Isolated polypeptide includes the polypeptide in situ within recombinant cells since at least one component of the polypeptide's natural environment will not be present. Ordinarily, however, isolated polypeptide will be prepared by at least one purification step. [0056]
  • “Treatment” refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented. [0057]
  • A “disorder” is any condition that would benefit from treatment with the polypeptide variant. This includes chronic and acute disorders or diseases including those pathological conditions which predispose the mammal to the disorder in question. [0058]
  • The word “label” when used herein refers to a detectable compound or composition which is conjugated directly or indirectly to the polypeptide. The label may be itself be detectable (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which is detectable. [0059]
  • An “isolated” nucleic acid molecule is a nucleic acid molecule that is identified and separated from at least one contaminant nucleic acid molecule with which it is ordinarily associated in the natural source of the polypeptide nucleic acid. An isolated nucleic acid molecule is other than in the form or setting in which it is found in nature. Isolated nucleic acid molecules therefore are distinguished from the nucleic acid molecule as it exists in natural cells. However, an isolated nucleic acid molecule includes a nucleic acid molecule contained in cells that ordinarily express the polypeptide where, for example, the nucleic acid molecule is in a chromosomal location different from that of natural cells. [0060]
  • The expression “control sequences” refers to DNA sequences necessary for the expression of an operably linked coding sequence in a particular host organism. The control sequences that are suitable for prokaryotes, for example, include a promoter, optionally an operator sequence, and a ribosome binding site. Eukaryotic cells are known to utilize promoters, polyadenylation signals, and enhancers. [0061]
  • Nucleic acid is “operably linked” when it is placed into a functional relationship with another nucleic acid sequence. For example, DNA for a presequence or secretory leader is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation. Generally, “operably linked” means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading phase. However, enhancers do not have to be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, the synthetic oligonucleotide adaptors or linkers are used in accordance with conventional practice. [0062]
  • As used herein, the expressions “cell,” “cell line,” and “cell culture” are used interchangeably and all such designations include progeny. Thus, the words “transformants” and “transformed cells” include the primary subject cell and cultures derived therefrom without regard for the number of transfers. It is also understood that all progeny may not be precisely identical in DNA content, due to deliberate or inadvertent mutations. Mutant progeny that have the same function or biological activity as screened for in the originally transformed cell are included. Where distinct designations are intended, it will be clear from the context. [0063]
  • The term “molecular complex” when used herein refers to the relatively stable structure which forms when two or more heterologous molecules (e.g. polypeptides) bind (preferably noncovalently) to one another. The preferred molecular complex herein is an immune complex. “Immune complex” refers to the relatively stable structure which forms when at least one target molecule and at least one heterologous Fc region-containing polypeptide bind to one another forming a larger molecular weight complex. Examples of immune complexes are antigen-antibody aggregates and target molecule-immunoadhesin aggregates. The term “immune complex” as used herein, unless indicated otherwise, refers to an ex vivo complex (i.e. other than the form or setting in which it may be found in nature). However, the immune complex may be administered to a mammal, e.g. to evaluate clearance of the immune complex in the mammal. [0064]
  • The term “target molecule” refers to a molecule, usually a polypeptide, which is capable of being bound by a heterologous molecule and has one or more binding sites for the heterologous molecule. The term “binding site” refers to a region of a molecule to which another molecule can bind. The “first target molecule” herein comprises at least two distinct binding sites (for example, two to five separate binding sites) for an analyte (e.g. an Fc region-containing polypeptide) such that at least two analyte molecules can bind to the first target molecule. In the preferred embodiment of the invention, the two or more binding sites are identical (e.g. having the same amino acid sequence, where the target molecule is a polypeptide). In Example 1 below, the first target molecule was IgE and had two separate binding sites in the Fc region thereof to which the Fc region-containing polypeptide (an anti-IgE antibody, E27) could bind. Other first target molecules include dimers of substantially identical monomors (e.g. neurotrophins, IL8 and VEGF) or are polypeptides comprising two or more substantially identical polypeptide chains (e.g. antibodies or immunoadhesins). The “second target molecule” comprises at least two distinct binding sites (for example, two to five separate binding sites) for the first target molecule such that at least two first target molecules can bind to the second target molecule. Preferably, the two or more binding sites are identical (e.g. having the same amino acid sequence, where the target molecule is a polypeptide). In Example 2, the second target molecule was VEGF, which has a pair of distinct binding sites to which the variable domain of the IgE antibody could bind. Other second target molecules are contemplated, e.g. other dimers of substantially identical monomers (e.g. neurotrophins or IL8) or polypeptides comprising two or more substantially identical domains (e.g. antibodies or immunoadhesins). [0065]
  • An “analyte” is a substance that is to be analyzed. The preferred analyte is an Fc region-containing polypeptide that is to be analyzed for its ability to bind to an Fc receptor. [0066]
  • A “receptor” is a polypeptide capable of binding at least one ligand. The preferred receptor is a cell-surface receptor having an extracellular ligand-binding domain and, optionally, other domains (e.g. transmembrane domain, intracellular domain and/or membrane anchor). The receptor to be evaluated in the assay described herein may be an intact receptor or a fragment or derivative thereof (e.g. a fusion protein comprising the binding domain of the receptor fused to one or more heterologous polypeptides). Moreover, the receptor to be evaluated for its binding properties may be present in a cell or isolated and optionally coated on an assay plate or some other solid phase. [0067]
  • The phrase “low affinity receptor” denotes a receptor that has a weak binding affinity for a ligand of interest, e.g. having a binding constant of about 50 nM or worse affinity. Exemplary low affinity receptors include FcγRII and FcγRIII as well as adhesion molecules, such as integrins. [0068]
  • II. Modes for Carrying out the Invention [0069]
  • The invention herein relates to a method for making a polypeptide variant. The “starting” or “nonvariant” polypeptide is prepared using techniques available in the art for generating polypeptides comprising an Fc region. In the preferred embodiment of the invention, the polypeptide is an antibody and exemplary methods for generating antibodies are described in more detail in the following sections. The polypeptide may, however, be any other polypeptide comprising an Fc region, e.g. an immunoadhesin. Methods for making immunoadhesins are elaborated in more detail hereinbelow. [0070]
  • The starting polypeptide of particular interest herein is usually one that binds to C1q and displays complement dependent cytotoxicity (CDC). The amino acid substitutions described herein will generally serve to alter the ability of the starting polypeptide to bind to C1q and/or modify its complement dependent cytotoxicity function, e.g. to reduce and preferably abolish these effector functions. However, polypeptides comprising substitutions at one or more of the described positions with improved effector functions are contemplated herein. For example, the starting polypeptide may be unable to bind C1q and/or mediate CDC and may be modified according to the teachings herein such that it acquires these effector functions. Moreover, polypeptides with pre-existing C1q binding activity, optionally further having the ability to mediate CDC may be modified such that one or both of these activities are enhanced. [0071]
  • To generate the polypeptide variant, one or more amino acid alterations (e.g. substitutions) are introduced in the Fc region of the starting polypeptide. The amino acid positions to be modified are selected from heavy chain positions 329, 331 and 322, where the numbering of the residues in an IgG heavy chain is that of the EU index as in Kabat et al, [0072] Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md (1991). The Fc region is preferably from a human IgG and most preferably a human IgG1. The human IgG1 Fc region may be a human A or non-A allotype. Proline is conserved at position 329 in human IgG's. This residue is preferably replaced with alanine, however substitution with any other amino acid is contemplated, e.g., serine, threonine, asparagine, glycine or valine. Proline is conserved at position 331 in human IgG1, IgG2 and IgG3, but not IgG4 (which has a serine residue at position 331). Residue 331 is preferably replaced by alanine or another amino acid, e.g. serine (for IgG regions other than IgG4), glycine or valine. Lysine 322 is conserved in human IgGs, and this residue is preferably replaced by an alanine residue, but substitution with any other amino acid residue is contemplated, e.g. serine, threonine, glycine or valine.
  • In one embodiment, only one of the three above-identified positions is altered in order to generate the polypeptide variant. Preferably only residue 329 or only residue 322 is altered if this is the case. Alternatively, two or all of the three identified positions are modified. Thus, a variant may be generated in which the native amino acid residue at position 329 of the human heavy chain constant region is substituted with another amino acid, optionally in combination with a substitution of the amino acid residue at position 331 and/or substitution of the amino acid residue at position 322. Otherwise, the native amino acid residue at position 331 and the native amino acid residue at position 322 of the human IgG Fc region may both be substituted with another amino acid residue. [0073]
  • DNA encoding amino acid sequence variant of the starting polypeptide is prepared by a variety of methods known in the art. These methods include, but are not limited to, preparation by site-directed (or oligonucleotide-mediated) mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlier prepared DNA encoding the polypeptide [0074]
  • Site-directed mutagenesis is a preferred method for preparing substitution variants. This technique is well known in the art (see, e.g.,Carter et al. [0075] Nucleic Acids Res. 13:4431-4443 (1985) and Kunkel et al, Proc. Natl. Acad.Sci.USA 82:488 (1987)). Briefly, in carrying out site-directed mutagenesis of DNA, the starting DNA is altered by first hybridizing an oligonucleotide encoding the desired mutation to a single strand of such starting DNA. After hybridization, a DNA polymerase is used to synthesize an entire second strand, using the hybridized oligonucleotide as a primer, and using the single strand of the starting DNA as a template. Thus, the oligonucleotide encoding the desired mutation is incorporated in the resulting double-stranded DNA.
  • PCR mutagenesis is also suitable for making amino acid sequence variants of the starting polypeptide. See Higuchi, in [0076] PCR Protocols, pp. 177-183 (Academic Press, 1990); and Vallette et al., Nuc. Acids Res. 17:723-733 (1989). Briefly, when small amounts of template DNA are used as starting material in a PCR, primers that differ slightly in sequence from the corresponding region in a template DNA can be used to generate relatively large quantities of a specific DNA fragment that differs from the template sequence only at the positions where the primers differ from the template.
  • Another method for preparing variants, cassette mutagenesis, is based on the technique described by Wells et al., [0077] Gene 34:315-323 (1985). The starting material is the plasmid (or other vector) comprising the starting polypeptide DNA to be mutated. The codon(s) in the starting DNA to be mutated are identified. There must be a unique restriction endonuclease site on each side of the identified mutation site(s). If no such restriction sites exist, they may be generated using the above-described oligonucleotide-mediated mutagenesis method to introduce them at appropriate locations in the starting polypeptide DNA. The plasmid DNA is cut at these sites to linearize it. A double-stranded oligonucleotide encoding the sequence of the DNA between the restriction sites but containing the desired mutation(s) is synthesized using standard procedures, wherein the two strands of the oligonucleotide are synthesized separately and then hybridized together using standard techniques. This double-stranded oligonucleotide is referred to as the cassette. This cassette is designed to have 5′ and 3′ ends that are compatible with the ends of the linearized plasmid, such that it can be directly ligated to the plasmid. This plasmid now contains the mutated DNA sequence.
  • The polypeptide variant(s) so prepared may be subjected to further modifications, oftentimes depending on the intended use of the polypeptide. Such modifications may involve further alteration of the amino acid sequence, fusion to heterologous polypeptide(s) and/or covalent modifications. For example, it may be useful to combine the above amino acid substitutions with one or more further amino acid substitutions that reduce or ablate FcR binding. For example, the native amino acid residues at any one or more of heavy chain positions 233-238, 318 or 331 (where the numbering of the residues in an IgG heavy chain is that of the EU index as in Kabat et al, supra) may be replaced with non-native residue(s), e.g. alanine. [0078]
  • With respect to further amino acid sequence alterations, any cysteine residue not involved in maintaining the proper conformation of the polypeptide variant also may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant cross linking. [0079]
  • Another type of amino acid substitution serves to alter the glycosylation pattern of the polypeptide. This may be achieved by deleting one or more carbohydrate moieties found in the polypeptide, and/or adding one or more glycosylation sites that are not present in the polypeptide. Glycosylation of polypeptides is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Thus, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N-aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used. Addition of glycosylation sites to the polypeptide is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites). The alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original polypeptide (for O-linked glycosylation sites). An exemplary glycosylation variant has an amino acid substitution of residue Asn 297 of the heavy chain. [0080]
  • The polypeptide variant may be subjected to one or more assays to evaluate any change in biological activity compared to the starting polypeptide. For example, the ability of the variant to bind C1q and mediate complement dependent cytotoxicity (CDC) may be assessed. [0081]
  • To determine C1q binding, a C1q binding ELISA may be performed. Briefly, assay plates may be coated overnight at 4° C. with polypeptide variant or starting polypeptide (control) in coating buffer. The plates may then be washed and blocked. Following washing, an aliquot of human C1q may be added to each well and incubated for 2 hrs at room temperature. Following a further wash, 100 μl of a sheep anti-complement C1q peroxidase conjugated antibody may be added to each well and incubated for 1 hour at room temperature. The plate may again be washed with wash buffer and 100 μl of substrate buffer containing OPD ( O-phenylenediamine dihydrochloride (Sigma)) may be added to each well. The oxidation reaction, observed by the appearance of a yellow color, may be allowed to proceed for 30 minutes and stopped by the addition of 100 μl of 4.5 N H[0082] 2SO4. The absorbance may then read at (492-405) nm.
  • The preferred polypeptide variant is one that displays a “significant reduction in C1q binding” in this assay. This means that about 100 μg/ml of the polypeptide variant displays about 50 fold or more reduction in C1q binding compared to 100 μg/ml of a control antibody having a nonmutated IgG1 Fc region. In the most preferred embodiment, the polypeptide variant “does not bind C1q”, i.e. 100 μg/ml of the polypeptide variant displays about 100 fold or more reduction in C1q binding compared to 100 μg/ml of the control antibody. [0083]
  • To assess complement activation, a complement dependent cytotoxicity (CDC) assay may be performed, e.g. as described in Gazzano-Santoro et al., [0084] J. Immunol. Methods 202:163 (1996). Briefly, various concentrations of the polypeptide variant and human complement may be diluted with buffer. Cells which express the antigen to which the polypeptide variant binds may be diluted to a density of˜1×106 cells/ ml. Mixtures of polypeptide variant, diluted human complement and cells expressing the antigen may be added to a flat bottom tissue culture 96 well plate and allowed to incubate for 2 hrs at 37°C. and 5% CO2 to facilitate complement mediated cell lysis. 50 μl of alamar blue (Accumed International) may then be added to each well and incubated overnight at 37°C. The absorbance is measured using a 96-well fluorometer with excitation at 530 nm and emission at 590 nm. The results may be expressed in relative fluorescence units (RFU). The sample concentrations may be computed from a standard curve and the percent activity as compared to nonvariant polypeptide is reported for the polypeptide variant of interest.
  • The preferred variant “does not activate complement”. For example, 0.6 μ/ml of the polypeptide variant displays about 0-10% CDC activity in this assay compared to a 0.6 μ/ml of a control antibody having a nonmutated IgG1 Fc region. Preferably the variant does not appear to have any CDC activity in the above CDC assay. [0085]
  • Preferably the polypeptide variant essentially retains the ability to bind antigen compared to the nonvariant polypeptide, i.e. the binding capability is no worse than about 20 fold, e.g. no worse than about 5 fold of that of the nonvariant polypeptide. The binding capability of the polypeptide variant may be determined using techniques such as fluorescence activated cell sorting (FACS) analysis or radioimmunoprecipitation (RIA), for example. [0086]
  • The ability of the polypeptide variant to bind an FcR may also be evaluated. Where the FcR is a high affinity Fc receptor, such as FcγRI or FcRn, binding can be measured by titrating monomeric polypeptide variant and measuring bound polypeptide variant using an antibody which specifically binds to the polypeptide variant in a standard ELISA format (see Example 2 below). Another FcR binding assay for low affinity FcRs is elaborated in more detail in the following section. [0087]
  • Preferably the variant retains the ability to bind one or more FcRs, e.g. the ability of the polypeptide variant to bind FcγRI, FcγRII, FcγRIII and/or FcRn is no more than about 20 fold reduced, preferably no more than about 10 fold reduced, and most preferably no more than about two fold reduced compared to the starting polypeptide as determined in the FcγRI or FcRn assays of Example 2 or the FcγRII or FcγRIII assays described in the following section. [0088]
  • A. Receptor Binding Assay and Immune Complex [0089]
  • A receptor binding assay has been developed herein which is particularly useful for determining binding of an analyte of interest to a receptor where the affinity of the analyte for the receptor is relatively weak, e.g. in the micromolar range as is the case for FcγRIIa, FcγRIIb, FcγRIIIa and FcγRIIIb. The method involves the formation of a molecular complex that has an improved avidity for the receptor of interest compared to the noncomplexed analyte. The preferred molecular complex is an immune complex comprising: (a) an Fc region-containing polypeptide (such as an antibody or an immunoadhesin); (b) a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide; and (c) a second target molecule which comprises at least two binding sites for the first target molecule. [0090]
  • In Example 1 below, the Fc region-containing polypeptide is an anti-IgE antibody, such as the E27 antibody (FIGS. [0091] 4A-4B). E27, when mixed with human IgE at an 1:1 molar ratio, forms a stable hexamer consisting of three E27 molecules and three IgE molecules. In Example 1 below, the “first target molecule” is a chimeric form of IgE in which the Fab portion of an anti-VEGF antibody is fused to the human IgE Fc portion and the “second target molecule” is the antigen to which the Fab binds (i.e. VEGF). Each molecule of IgE binds two molecules of VEGF. VEGF also binds two molecules of IgE per molecule of VEGF. When recombinant human VEGF was added at a 2:1 molar ratio to IgE:E27 hexamers, the hexamers were linked into larger molecular weight complexes via the IgE:VEGF interaction (FIG. 5). The Fc region of the anti-IgE antibody of the resultant immune complex binds to FcR with higher avidity than either uncomplexed anti-IgE or anti-IgE:lgE hexamers.
  • Other forms of molecular complexes for use in the receptor assay are contemplated. Examples comprising only an Fc region-containing polypeptide:first target molecule combination include an immunoadhesin:ligand combination such as VEGF receptor (KDR)-immunoadhesin: VEGF and a full-length bispecific antibody (bsAb):first target molecule. A further example of an Fc region-containing polypeptide:first target molecule:second target molecule combination include a nonblocking antibody:soluble receptor:ligand combination such as anti-Trk antibody:soluble Trk receptor:neurotrophin (Urfer et al. [0092] J. Biol Chem. 273(10):5829-5840 (1998)).
  • Aside from use in a receptor binding assay, the immune complexes described above have further uses including evaluation of Fc region-containing polypeptide function and immune complex clearance in vivo. Hence, the immune complex may be administered to a mammal (e.g. in a pre-clinical animal study) and evaluated for its half-life etc. [0093]
  • To determine receptor binding, a polypeptide comprising at least the binding domain of the receptor of interest (e.g. the extracellular domain of an α subunit of an FcR) may be coated on solid phase, such as an assay plate. The binding domain of the receptor alone or a receptor-fusion protein may be coated on the plate using standard procedures. Examples of receptor-fusion proteins include receptor-glutathione S-transferase (GST) fusion protein, receptor-chitin binding domain fusion protein, receptor-hexaHis tag fusion protein (coated on glutathione, chitin, and nickel coated plates, respectively). Alternatively, a capture molecule may be coated on the assay plate and used to bind the receptor-fusion protein via the non-receptor portion of the fusion protein. Examples include anti-hexaHis F(ab′)[0094] 2 coated on the assay plate used to capture receptor-hexaHis tail fusion or anti-GST antibody coated on the assay plate used to capture a receptor-GST fusion. In other embodiments, binding to cells expressing at least the binding domain of the receptor may be evaluated. The cells may be naturally occurring hematopoietic cells that express the FcR of interest or may be transformed with nucleic acid encoding the FcR or a binding domain thereof such that the binding domain is expressed at the surface of the cell to be tested.
  • The immune complex described hereinabove is added to the receptor-coated plates and incubated for a sufficient period of time such that the analyte binds to the receptor. Plates may then be washed to remove unbound complexes, and binding of the analyte may be detected according to known methods. For example, binding may be detected using a reagent (e.g. an antibody or fragment thereof) which binds specifically to the analyte, and which is optionally conjugated with a detectable label (detectable labels and methods for conjugating them to polypeptides are described below in the section entitled “Non-Therapeutic Uses for the Polypeptide Variant”). [0095]
  • As a matter of convenience, the reagents can be provided in an assay kit, i.e., a packaged combination of reagents, for combination with the analyte in assaying the ability of the analyte to bind to a receptor of interest. The components of the kit will generally be provided in predetermined ratios. The kit may provide the first target molecule and/or the second target molecule, optionally complexed together. The kit may further include assay plates coated with the receptor or a binding domain thereof (e.g. the extracellular domain of the αsubunit of an FcR). Usually, other reagents, such as an antibody that binds specifically to the analyte to be assayed, labeled directly or indirectly with an enzymatic label, will also be provided in the kit. Where the detectable label is an enzyme, the kit will include substrates and cofactors required by the enzyme (e.g. a substrate precursor which provides the detectable chromophore or fluorophore). In addition, other additives may be included such as stabilizers, buffers (e.g. assay and/or wash lysis buffer) and the like. The relative amounts of the various reagents may be varied widely to provide for concentrations in solution of the reagents that substantially optimize the sensitivity of the assay. Particularly, the reagents may be provided as dry powders, usually lyophilized, including excipients that on dissolution will provide a reagent solution having the appropriate concentration. The kit also suitably includes instructions for carrying out the assay. [0096]
  • B. Antibody Preparation [0097]
  • In the preferred embodiment of the invention, the Fc region-containing polypeptide which is modified according to the teachings herein is an antibody. Techniques for producing antibodies follow: [0098]
  • (i) Antigen Selection and Preparation [0099]
  • Where the polypeptide is an antibody, it is directed against an antigen of interest. Preferably, the antigen is a biologically important polypeptide and administration of the antibody to a mammal suffering from a disease or disorder can result in a therapeutic benefit in that mammal. However, antibodies directed against nonpolypeptide antigens (such as tumor-associated glycolipid antigens; see US Patent 5,091,178) are also contemplated. [0100]
  • Where the antigen is a polypeptide, it may be a transmembrane molecule (e.g. receptor) or ligand such as a growth factor. Exemplary antigens include molecules such as renin; a growth hormone, including human growth hormone and bovine growth hormone; growth hormone releasing factor; parathyroid hormone; thyroid stimulating hormone; lipoproteins; alpha-1-antitrypsin; insulin A-chain; insulin B-chain; proinsulin; follicle stimulating hormone; calcitonin; luteinizing hormone; glucagon; clofting factors such as factor VIIIC, factor IX, tissue factor, and von Willebrands factor; anti-clofting factors such as Protein C; atrial natriuretic factor; lung surfactant; a plasminogen activator, such as urokinase or human urine or tissue-type plasminogen activator (t-PA); bombesin; thrombin; hemopoietic growth factor; tumor necrosis factor-alpha and -beta; enkephalinase; RANTES (regulated on activation normally T-cell expressed and secreted); human macrophage inflammatory protein (MlP-1-alpha); a serum albumin such as human serum albumin; Muellerian-inhibiting substance; relaxin A-chain; relaxin B-chain; prorelaxin; mouse gonadotropin-associated peptide; a microbial protein, such as beta-lactamase; DNase; IgE; a cytotoxic T-lymphocyte associated antigen (CTLA), such as CTLA-4; inhibin; activin; vascular endothelial growth factor (VEGF); receptors for hormones or growth factors; protein A or D; rheumatoid factors; a neurotrophic factor such as bone-derived neurotrophic factor (BDNF), neurotrophin-3, -4, -5, or -6 (NT-3, NT-4, NT-5, or NT-6), or a nerve growth factor such as NGF-β; platelet-derived growth factor (PDGF); fibroblast growth factor such as aFGF and bFGF; epidermal growth factor (EGF); transforming growth factor (TGF) such as TGF-alpha and TGF-beta, including TGF-,β1, TGF-,β2, TGF-β3, TGF-β4, or TGF-β5; insulin-like growth factor-I and -II (IGF-I and IGF-II); des(1-3)-IGF-I (brain IGF-I), insulin-like growth factor binding proteins; CD proteins such as CD3, CD4, CD8, CD19 and CD20; erythropoietin; osteoinductive factors; immunotoxins; a bone morphogenetic protein (BMP); an interferon such as interferon-alpha, -beta, and -gamma; colony stimulating factors (CSFs), e.g., M-CSF, GM-CSF, and G-CSF; interieukins (ILs), e.g., IL-1 to IL-10; superoxide dismutase; T-cell receptors; surface membrane proteins; decay accelerating factor; viral antigen such as, for example, a portion of the AIDS envelope; transport proteins; homing receptors; addressins; regulatory proteins; integrins such as CD11a, CD11b, CD11c, CD18, an ICAM, VLA-4 and VCAM; a tumor associated antigen such as HER2, HER3 or HER4 receptor; and fragments of any of the above-listed polypeptides. [0101]
  • Preferred molecular targets for antibodies encompassed by the present invention include CD proteins such as CD3, CD4, CD8, CD19, CD20 and CD34; members of the ErbB receptor family such as the EGF receptor, HER2, HER3 or HER4 receptor; cell adhesion molecules such as LFA-1, Mac1, p150.95, VLA-4, ICAM-1, VCAM and αv/β3 integrin including either α or β subunits thereof (e.g. anti-CD11[0102] a, anti-CD18 or anti-CD11 b antibodies); growth factors such as VEGF; IgE; blood group antigens; flk2/flt3 receptor; obesity (OB) receptor; mpl receptor; CTLA-4; protein C etc.
  • Soluble antigens or fragments thereof, optionally conjugated to other molecules, can be used as immunogens for generating antibodies. For transmembrane molecules, such as receptors, fragments of these (e.g. the extracellular domain of a receptor) can be used as the immunogen. Alternatively, cells expressing the transmembrane molecule can be used as the immunogen. Such cells can be derived from a natural source (e.g. cancer cell lines) or may be cells which have been transformed by recombinant techniques to express the transmembrane molecule. Other antigens and forms thereof useful for preparing antibodies will be apparent to those in the art. [0103]
  • (ii) Polyclonal antibodies [0104]
  • Polyclonal antibodies are preferably raised in animals by multiple subcutaneous (sc) or intraperitoneal (ip) injections of the relevant antigen and an adjuvant. It may be useful to conjugate the relevant antigen to a protein that is immunogenic in the species to be immunized, e.g., keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor using a bifunctional or derivatizing agent, for example, maleimidobenzoyl sulfosuccinimide ester (conjugation through cysteine residues), N-hydroxysuccinimide (through lysine residues), glutaraldehyde, succinic anhydride, SOCl[0105] 2, or R1N═C═NR, where R and R1 are different alkyl groups.
  • Animals are immunized against the antigen, immunogenic conjugates, or derivatives by combining, e.g., 100 μg or 5 μg of the protein or conjugate (for rabbits or mice, respectively) with 3 volumes of Freund's complete adjuvant and injecting the solution intradermally at multiple sites. One month later the animals are boosted with 1/5 to 1/10 the original amount of peptide or conjugate in Freund's complete adjuvant by subcutaneous injection at multiple sites. Seven to 14 days later the animals are bled and the serum is assayed for antibody titer. Animals are boosted until the titer plateaus. Preferably, the animal is boosted with the conjugate of the same antigen, but conjugated to a different protein and/or through a different cross-linking reagent. Conjugates also can be made in recombinant cell culture as protein fusions. Also, aggregating agents such as alum are suitably used to enhance the immune response. [0106]
  • (iii) Monoclonal antibodies [0107]
  • Monoclonal antibodies may be made using the hybridoma method first described by Kohler et al, Nature, 256:495 (1975), or may be made by recombinant DNA methods (U.S. Pat. No. 4,816,567). [0108]
  • In the hybridoma method, a mouse or other appropriate host animal, such as a hamster or macaque monkey, is immunized as hereinabove described to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization. Alternatively, lymphocytes may be immunized in vitro. Lymphocytes then are fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, [0109] Monoclonal Antibodies: Principles and Practice, pp.59-103 (Academic Press, 1986)).
  • The hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells. For example, if the parental myeloma cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells. [0110]
  • Preferred myeloma cells are those that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. Among these, preferred myeloma cell lines are murine myeloma lines, such as those derived from MOPC-21 and MPC-11 mouse tumors available from the Salk Institute Cell Distribution Center, San Diego, Calif. USA, and SP-2 or X63-Ag8-653 cells available from the American Type Culture Collection, Rockville, Md. USA. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, [0111] J. Immunol., 133:3001 (1984); Brodeur et al, Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).
  • Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA). [0112]
  • After hybridoma cells are identified that produce antibodies of the desired specificity, affinity, and/or activity, the clones may be subcloned by limiting dilution procedures and grown by standard methods (Goding, [0113] Monoclonal Antibodies: Principles and Practice, pp.59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium. In addition, the hybridoma cells may be grown in vivo as ascites tumors in an animal.
  • The monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography. [0114]
  • DNA encoding the monoclonal antibodies is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the monoclonal antibodies). The hybridoma cells serve as a preferred source of such DNA. Once isolated, the DNA may be placed into expression vectors, which are then transfected into host cells such as [0115] E. coi cells, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. Recombinant production of antibodies will be described in more detail below.
  • In a further embodiment, antibodies or antibody fragments can be isolated from antibody phage libraries generated using the techniques described in McCafferty et al, [0116] Nature, 348:552-554 1990). Clackson et al., Nature, 352:624-628 (1991) and Marks et al, J. Mol. Biol., 222:581-597 (1991) describe the isolation of murine and human antibodies, respectively, using phage libraries. Subsequent publications describe the production of high affinity (nM range) human antibodies by chain shuffling (Marks et al, Bio/Technology, 10:779-783 (1992)), as well as combinatorial infection and in vivo recombination as a strategy for constructing very large phage libraries (Waterhouse et al., Nuc. Acids. Res., 21:2265-2266 (1993)). Thus, these techniques are viable alternatives to traditional monoclonal antibody hybridoma techniques for isolation of monoclonal antibodies.
  • The DNA also may be modified, for example, by substituting the coding sequence for human heavy- and light-chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567; Morrison, et al., [0117] Proc. Natl Acad. Sci. USA, 81:6851 (1984)), or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide.
  • Typically such non-immunoglobulin polypeptides are substituted for the constant domains of an antibody, or they are substituted for the variable domains of one antigen-combining site of an antibody to create a chimeric bivalent antibody comprising one antigen-combining site having specificity for an antigen and another antigen-combining site having specificity for a different antigen. [0118]
  • (iv) Humanized and Human Antibodies [0119]
  • A humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as “import” residues, which are typically taken from an “import” variable domain. Humanization can be essentially performed following the method of Winter and co-workers (Jones et al., Nature, 321:522-525 (1986); Riechmann et al, [0120] Nature, 332:323-327 (1988); Verhoeyen et al, Science, 239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Accordingly, such “humanized” antibodies are chimeric antibodies (U.S. Pat. No. 4,816,567) wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
  • The choice of human variable domains, both light and heavy, to be used in making the humanized antibodies is very important to reduce antigenicity. According to the so-called “best-fit” method, the sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable-domain sequences. The human sequence which is closest to that of the rodent is then accepted as the human framework (FR) for the humanized antibody (Sims et al, [0121] J. Immunol., 151:2296 (1993); Chothia et al., J. Mol Biol, 196:901 (1987)). Another method uses a particular framework derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains. The same framework may be used for several different humanized antibodies (Carter et al., Proc. Natl. Acad. Sci. USA, 89:4285 (1992); Presta et al., J. Immnol., 151:2623 (1993)).
  • It is further important that antibodies be humanized with retention of high affinity for the antigen and other favorable biological properties. To achieve this goal, according to a preferred method, humanized antibodies are prepared by a process of analysis of the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences. Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen. In this way, FR residues can be selected and combined from the recipient and import sequences so that the desired antibody characteristic, such as increased affinity for the target antigen(s), is achieved. In general, the CDR residues are directly and most substantially involved in influencing antigen binding. [0122]
  • Alternatively, it is now possible to produce transgenic animals (e.g., mice) that are capable, upon immunization, of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production. For example, it has been described that the homozygous deletion of the antibody heavy-chain joining region (J[0123] H) gene in chimeric and germ-line mutant mice results in complete inhibition of endogenous antibody production. Transfer of the human germ-line immunoglobulin gene array in such germ-line mutant mice will result in the production of human antibodies upon antigen challenge. See, e.g., Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggermann et al, Year in Immuno., 7:33 (1993); and Duchosal et al. Nature 355:258 (1992). Human antibodies can also be derived from phage-display libraries (Hoogenboom et al., J. MoL Biol., 227:381 (1991); Marks et al., J. Mol. Biol, 222:581-597 (1991); Vaughan et al. Nature Biotech 14:309 (1996)).
  • (v) Multispecific Antibodies [0124]
  • Multispecific antibodies have binding specificities for at least two different antigens. While such molecules normally will only bind two antigens (i.e. bispecific antibodies, BsAbs), antibodies with additional specificities such as trispecific antibodies are encompassed by this expression when used herein. Examples of BsAbs include those with one arm directed against a tumor cell antigen and the other arm directed against a cytotoxic trigger molecule such as anti-FcγRl/anti-CD15, anti-p185[0125] HER2/FcγRIII (CD16), anti-CD3/anti-malignant B-cell (1D10), anti-CD3/anti-p185HER2, anti-CD3/anti-p97, anti-CD3/anti-renal cell carcinoma, anti-CD3/anti-OVCAR-3, anti-CD3/L-D1 (anti-colon carcinoma), anti-CD3/anti-melanocyte stimulating hormone analog, anti-EGF receptor/anti-CD3, anti-CD3/anti-CAMA1, anti-CD3/anti-CD19, anti-CD3/MoV18, anti-neural cell ahesion molecule (NCAM)/anti-CD3, anti-folate binding protein (FBP)/anti-CD3, anti-pan carcinoma associated antigen (AMOC-31)/anti-CD3; BsAbs with one arm which binds specifically to a tumor antigen and one arm which binds to a toxin such as anti-saporin/anti-ld-1, anti-CD22/anti-saporin, anti-CD7/anti-saporin, anti-CD38/anti-saporin, anti-CEA/anti-ricin A chain, anti-interferon-α(IFN-α)/anti-hybridoma idiotype, anti-CEA/anti-vinca alkaloid; BsAbs for converting enzyme activated prodrugs such as anti-CD30/anti-alkaline phosphatase (which catalyzes conversion of mitomycin phosphate prodrug to mitomycin alcohol); BsAbs which can be used as fibrinolytic agents such as anti-fibrin/anti-tissue plasminogen activator (tPA), anti-fibrin/anti-urokinase-type plasminogen activator (uPA); BsAbs for targeting immune complexes to cell surface receptors such as anti-low density lipoprotein (LDL)/anti-Fc receptor (e.g. FcγRI, FcγRII or FcγRIII); BsAbs for use in therapy of infectious diseases such as anti-CD3/anti-herpes simplex virus (HSV), anti-T-cell receptor:CD3 complex/anti-influenza, anti-FcγR/anti-HIV; BsAbs for tumor detection in vitro or in vivo such as anti-CEA/anti-EOTUBE, anti-CEA/anti-DPTA, anti-p185 HER2/anti-hapten; BsAbs as vaccine adjuvants; and BsAbs as diagnostic tools such as anti-rabbit IgG/anti-ferritin, anti-horse radish peroxidase (HRP)/anti-hormone, anti-somatostatin/anti-substance P, anti-HRP/anti-FITC, anti-CEA/anti-β-galactosidase. Examples of trispecific antibodies include anti-CD3/anti-CD4/anti-CD37, anti-CD3/anti-CD5/anti-CD37 and anti-CD3/anti-CD8/anti-CD37. Bispecific antibodies can be prepared as full length antibodies or antibody fragments (e.g. F(ab′)2 bispecific antibodies).
  • Methods for making bispecific antibodies are known in the art. Traditional production of full length bispecific antibodies is based on the coexpression of two immunoglobulin heavy chain-light chain pairs, where the two chains have different specificities (Millstein et al., [0126] Nature, 305:537-539 (1983)). Because of the random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture of 10 different antibody molecules, of which only one has the correct bispecific structure. Purification of the correct molecule, which is usually done by affinity chromatography steps, is rather cumbersome, and the product yields are low. Similar procedures are disclosed in WO 93/08829, and in Traunecker et al., EMBO J., 10:3655-3659(1991).
  • According to a different approach, antibody variable domains with the desired binding specificities (antibody-antigen combining sites) are fused to immunoglobulin constant domain sequences. The fusion preferably is with an immunoglobulin heavy chain constant domain, comprising at least part of the hinge, CH2, and CH3 regions. It is preferred to have the first heavy-chain constant region (CH1) containing the site necessary for light chain binding, present in at least one of the fusions. DNAs encoding the immunoglobulin heavy chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co-transfected into a suitable host organism. This provides for great flexibility in adjusting the mutual proportions of the three polypeptide fragments in embodiments when unequal ratios of the three polypeptide chains used in the construction provide the optimum yields. It is, however, possible to insert the coding sequences for two or all three polypeptide chains in one expression vector when the expression of at least two polypeptide chains in equal ratios results in high yields or when the ratios are of no particular significance. [0127]
  • In a preferred embodiment of this approach, the bispecific antibodies are composed of a hybrid immunoglobulin heavy chain with a first binding specificity in one arm, and a hybrid immunoglobulin heavy chain-light chain pair (providing a second binding specificity) in the other arm. It was found that this asymmetric structure facilitates the separation of the desired bispecific compound from unwanted immunoglobulin chain combinations, as the presence of an immunoglobulin light chain in only one half of the bispecific molecule provides for a facile way of separation. This approach is disclosed in WO 94/04690. For further details of generating bispecific antibodies see, for example, Suresh et al., [0128] Methods in Enzymology, 121:210 (1986).
  • According to another approach described in W096/27011, the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers which are recovered from recombinant cell culture. The preferred interface comprises at least a part of the [0129] C H 3 domain of an antibody constant domain. In this method, one or more small amino acid side chains from the interface of the first antibody molecule are replaced with larger side chains (e.g. tyrosine or tryptophan). Compensatory “cavities” of identical or similar size to the large side chain(s) are created on the interface of the second antibody molecule by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine). This provides a mechanism for increasing the yield of the heterodimer over other unwanted end-products such as homodimers.
  • Bispecific antibodies include cross-linked or “heteroconjugate” antibodies. For example, one of the antibodies in the heteroconjugate can be coupled to avidin, the other to biotin. Such antibodies have, for example, been proposed to target immune system cells to unwanted cells (U.S. Pat. No. 4,676,980), and for treatment of HIV infection (WO 91/00360, WO 921200373, and EP 03089). Heteroconjugate antibodies may be made using any convenient cross-linking methods. Suitable cross-linking agents are well known in the art, and are disclosed in U.S. Pat. No. 4,676,980, along with a number of cross-linking techniques. [0130]
  • Antibodies with more than two valencies are contemplated. For example, trispecific antibodies can be prepared. Tutt et al [0131] J. Immunol. 147:60 (1991).
  • While the polypeptide of interest herein is preferably an antibody, other Fc region-containing polypeptides which can be modified according to the methods described herein are contemplated. An example of such a molecule is an immunoadhesin. [0132]
  • C. Immunoadhesin Preparation [0133]
  • The simplest and most straightforward immunoadhesin design combines the binding domain(s) of the adhesin (e.g. the extracellular domain (ECD) of a receptor) with the Fc region of an immunoglobulin heavy chain. Ordinarily, when preparing the immunoadhesins of the present invention, nucleic acid encoding the binding domain of the adhesin will be fused C-terminally to nucleic acid encoding the N-terminus of an immunoglobulin constant domain sequence, however N-terminal fusions are also possible. [0134]
  • Typically, in such fusions the encoded chimeric polypeptide will retain at least functionally active hinge, [0135] C H2 and C H3 domains of the constant region of an immunoglobulin heavy chain. Fusions are also made to the C-terminus of the Fc portion of a constant domain, or immediately N-terminal to the C H1 of the heavy chain or the corresponding region of the light chain. The precise site at which the fusion is made is not critical; particular sites are well known and may be selected in order to optimize the biological activity, secretion, or binding characteristics of the immunoadhesin.
  • In a preferred embodiment, the adhesin sequence is fused to the N-terminus of the Fc region of immunoglobulin G[0136] 1 (IgG1). It is possible to fuse the entire heavy chain constant region to the adhesin sequence. However, more preferably, a sequence beginning in the hinge region just upstream of the papain cleavage site which defines IgG Fc chemically (i.e. residue 216, taking the first residue of heavy chain constant region to be 114), or analogous sites of other immunoglobulins is used in the fusion. In a particularly preferred embodiment, the adhesin amino acid sequence is fused to (a) the hinge region and C H 2 and C H 3 or (b) the C H1, hinge, C H2 and C H3 domains, of an IgG heavy chain.
  • For bispecific immunoadhesins, the immunoadhesins are assembled as multimers, and particularly as heterodimers or heterotetramers. Generally, these assembled immunoglobulins will have known unit structures. A basic four chain structural unit is the form in which IgG, lgD, and IgE exist. A four chain unit is repeated in the higher molecular weight immunoglobulins; IgM generally exists as a pentamer of four basic units held together by disulfide bonds. IgA globulin, and occasionally IgG globulin, may also exist in multimeric form in serum. In the case of multimer, each of the four units may be the same or different. [0137]
  • Various exemplary assembled immunoadhesins within the scope herein are schematically diagrammed below: [0138]
  • (a) AC[0139] L-ACL;
  • (b) AC[0140] H-(ACH, ACL-ACH, ACL-VHCH, or VLCL-ACH);
  • (c) AC[0141] L-ACH-(ACL-ACH, ACL-VHCH, VLCL-ACH, or VLCL-VH CH)
  • (d) AC[0142] L-VHCH-(ACH, or ACL-VHCH, or VLCL-ACH);
  • (e) V[0143] LCL-ACH-(ACL-VHCH, or VLCL-ACH); and
  • (f) (A-Y)[0144] n-(VLCL-VHCH)2,
  • wherein each A represents identical or different adhesin amino acid sequences; [0145]
  • V[0146] L is an immunoglobulin light chain variable domain;
  • V[0147] H is an immunoglobulin heavy chain variable domain;
  • C[0148] L is an immunoglobulin light chain constant domain;
  • C[0149] H is an immunoglobulin heavy chain constant domain;
  • n is an integer greater than 1; [0150]
  • Y designates the residue of a covalent cross-linking agent. [0151]
  • In the interests of brevity, the foregoing structures only show key features; they do not indicate joining (J) or other domains of the immunoglobulins, nor are disulfide bonds shown. However, where such domains are required for binding activity, they shall be constructed to be present in the ordinary locations which they occupy in the immunoglobulin molecules. [0152]
  • Alternatively, the adhesin sequences can be inserted between immunoglobulin heavy chain and light chain sequences, such that an immunoglobulin comprising a chimeric heavy chain is obtained. In this embodiment, the adhesin sequences are fused to the 3′ end of an immunoglobulin heavy chain in each arm of an immunoglobulin, either between the hinge and the [0153] C H2 domain, or between the C H2 and C H3 domains. Similar constructs have been reported by Hoogenboom, et al., Mol. Immunol. 28:1027-1037 (1991).
  • Although the presence of an immunoglobulin light chain is not required in the immunoadhesins of the present invention, an immunoglobulin light chain might be present either covalently associated to an adhesin-immunoglobulin heavy chain fusion polypeptide, or directly fused to the adhesin. In the former case, DNA encoding an immunoglobulin light chain is typically coexpressed with the DNA encoding the adhesin-immunoglobulin heavy chain fusion protein. Upon secretion, the hybrid heavy chain and the light chain will be covalently associated to provide an immunoglobulin-like structure comprising two disulfide-linked immunoglobulin heavy chain-light chain pairs. Methods suitable for the preparation of such structures are, for example, disclosed in U.S. Pat. No. 4,816,567, issued Mar. 28, 1989. [0154]
  • lmmunoadhesins are most conveniently constructed by fusing the cDNA sequence encoding the adhesin portion in-frame to an immunoglobulin cDNA sequence. However, fusion to genomic immunoglobulin fragments can also be used (see, e.g. Aruffo et al, [0155] Cell61:1303-1313 (1990); and Stamenkovic et al, Cell 66:1133-1144 (1991)). The latter type of fusion requires the presence of Ig regulatory sequences for expression. cDNAs encoding IgG heavy-chain constant regions can be isolated based on published sequences from cDNA libraries derived from spleen or peripheral blood lymphocytes, by hybridization or by polymerase chain reaction (PCR) techniques. The cDNAs encoding the “adhesin” and the immunoglobulin parts of the immunoadhesin are inserted in tandem into a plasmid vector that directs efficient expression in the chosen host cells.
  • D. Vectors, Host Cells and Recombinant Methods [0156]
  • The invention also provides isolated nucleic acid encoding a polypeptide variant as disclosed herein, vectors and host cells comprising the nucleic acid, and recombinant techniques for the production of the polypeptide variant. [0157]
  • For recombinant production of the polypeptide variant, the nucleic acid encoding it is isolated and inserted into a replicable vector for further cloning (amplification of the DNA) or for expression. DNA encoding the polypeptide variant is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the polypeptide variant). Many vectors are available. The vector components generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence. [0158]
  • (i) Signal Sequence Component [0159]
  • The polypeptide variant of this invention may be produced recombinantly not only directly, but also as a fusion polypeptide with a heterologous polypeptide, which is preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide. The heterologous signal sequence selected preferably is one that is recognized and processed (ie., cleaved by a signal peptidase) by the host cell. For prokaryotic host cells that do not recognize and process the native polypeptide variant signal sequence, the signal sequence is substituted by a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II leaders. For yeast secretion the native signal sequence may be substituted by, e.g., the yeast invertase leader, α factor leader (including Saccharomyces and Kluyveromyces α-factor leaders), or acid phosphatase leader, the [0160] C. albicans glucoamylase leader, or the signal described in WO 90113646. In mammalian cell expression, mammalian signal sequences as well as viral secretory leaders, for example, the herpes simplex gD signal, are available.
  • The DNA for such precursor region is ligated in reading frame to DNA encoding the polypeptide variant. [0161]
  • (ii) Origin of Replication Component Both Expression and Cloning Vectors Contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells. Generally, in cloning vectors this sequence is one that enables the vector to replicate independently of the host chromosomal DNA, and includes origins of replication or autonomously replicating sequences. Such sequences are well known for a variety of bacteria, yeast, and viruses. The origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria, the 2μ plasmid origin is suitable for yeast, and various viral origins (SV40, polyoma, adenovirus, VSV or BPV) are useful for cloning vectors in mammalian cells. Generally, the origin of replication component is not needed for mammalian expression vectors (the SV40 origin may typically be used only because it contains the early promoter). [0162]
  • (iii) Selection gene component [0163]
  • Expression and cloning vectors may contain a selection gene, also termed a selectable marker. Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli. [0164]
  • One example of a selection scheme utilizes a drug to arrest growth of a host cell. Those cells that are successfully transformed with a heterologous gene produce a protein conferring drug resistance and thus survive the selection regimen. Examples of such dominant selection use the drugs neomycin, mycophenolic acid and hygromycin. [0165]
  • Another example of suitable selectable markers for mammalian cells are those that enable the identification of cells competent to take up the polypeptide variant nucleic acid, such as DHFR, thymidine kinase, metallothionein-I and -II, preferably primate metallothionein genes, adenosine deaminase, ornithine decarboxylase, etc. [0166]
  • For example, cells transformed with the DHFR selection gene are first identified by culturing all of the transformants in a culture medium that contains methotrexate (Mtx), a competitive antagonist of DHFR. An appropriate host cell when wild-type DHFR is employed is the Chinese hamster ovary (CHO) cell line deficient in DHFR activity. [0167]
  • Alternatively, host cells (particularly wild-type hosts that contain endogenous DHFR) transformed or co-transformed with DNA sequences encoding polypeptide variant, wild-type DHFR protein, and another selectable marker such as [0168] aminoglycoside 3′-phosphotransferase (APH) can be selected by cell growth in medium containing a selection agent for the selectable marker such as an aminoglycosidic antibiotic, e.g., kanamycin, neomycin, or G418. See U.S. Pat. No. 4,965,199.
  • A suitable selection gene for use in yeast is the trpl gene present in the yeast plasmid YRp7 (Stinchcomb et al., [0169] Nature, 282:39 (1979)). The trpl gene provides a selection marker for a mutant strain of yeast lacking the ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1. Jones, Genetics, 85:12 (1977). The presence of the trp1 lesion in the yeast host cell genome then provides an effective environment for detecting transformation by growth in the absence of tryptophan. Similarly, Leu2-deficient yeast strains (ATCC 20,622 or 38,626) are complemented by known plasmids bearing the Leu2 gene.
  • In addition, vectors derived from the 1.6 μm circular plasmid pKDl can be used for transformation of Kluyveromyces yeasts. Alternatively, an expression system for large-scale production of recombinant calf chymosin was reported for K. lactis. Van den Berg, [0170] Bio/Technology, 8:135 (1990). Stable multi-copy expression vectors for secretion of mature recombinant human serum albumin by industrial strains of Kluyveromyces have also been disclosed. Fleer et al., Bio/Technology, 9:968-975 (1991).
  • (iv) Promoter component [0171]
  • Expression and cloning vectors usually contain a promoter that is recognized by the host organism and is operably linked to the polypeptide variant nucleic acid. Promoters suitable for use with prokaryotic hosts include the phoA promoter, β-lactamase and lactose promoter systems, alkaline phosphatase, a tryptophan (trp) promoter system, and hybrid promoters such as the tac promoter. However, other known bacterial promoters are suitable. Promoters for use in bacterial systems also will contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA encoding the polypeptide variant. [0172]
  • Promoter sequences are known for eukaryotes. Virtually all eukaryotic genes have an AT-rich region located approximately 25 to 30 bases upstream from the site where transcription is initiated. Another sequence found 70 to 80 bases upstream from the start of transcription of many genes is a CNCAAT region where N may be any nucleotide. At the 3′ end of most eukaryotic genes is an AATAAA sequence that may be the signal for addition of the poly A tail to the 3′ end of the coding sequence. All of these sequences are suitably inserted into eukaryotic expression vectors. [0173]
  • Examples of suitable promoting sequences for use with yeast hosts include the promoters for 3-phosphoglycerate kinase or other glycolytic enzymes, such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate kinase, triosephosphate isomerase, phosphoglucose isomerase, and glucokinase. [0174]
  • Other yeast promoters, which are inducible promoters having the additional advantage of transcription controlled by growth conditions, are the promoter regions for [0175] alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, degradative enzymes associated with nitrogen metabolism, metallothionein, glyceraldehyde-3-phosphate dehydrogenase, and enzymes responsible for maltose and galactose utilization. Suitable vectors and promoters for use in yeast expression are further described in EP 73,657. Yeast enhancers also are advantageously used with yeast promoters.
  • Polypeptide variant transcription from vectors in mammalian host cells is controlled, for example, by promoters obtained from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and most preferably Simian Virus 40 (SV40), from heterologous mammalian promoters, e.g., the actin promoter or an immunoglobulin promoter, from heat-shock promoters, provided such promoters are compatible with the host cell systems. [0176]
  • The early and late promoters of the SV40 virus are conveniently obtained as an SV40 restriction fragment that also contains the SV40 viral origin of replication. The immediate early promoter of the human cytomegalovirus is conveniently obtained as a HindIII E restriction fragment. A system for expressing DNA in mammalian hosts using the bovine papilloma virus as a vector is disclosed in U.S. Pat. No. 4,419,446. A modification of this system is described in U.S. Pat. No. 4,601,978. See also Reyes et al, [0177] Nature 297:598-601 (1982) on expression of human β-interferon cDNA in mouse cells under the control of a thymidine kinase promoter from herpes simplex virus. Alternatively, the rous sarcoma virus long terminal repeat can be used as the promoter.
  • (v) Enhancer Element Component [0178]
  • Transcription of a DNA encoding the polypeptide variant of this invention by higher eukaryotes is often increased by inserting an enhancer sequence into the vector. Many enhancer sequences are now known from mammalian genes (globin, elastase, albumin, α-fetoprotein, and insulin). Typically, however, one will use an enhancer from a eukaryotic cell virus. Examples include the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers. See also Yaniv, [0179] Nature 297:17-18 (1982) on enhancing elements for activation of eukaryotic promoters. The enhancer may be spliced into the vector at a position 5′ or 3′ to the polypeptide variant-encoding sequence, but is preferably located at a site 5′ from the promoter.
  • (vi) Transcription Termination Component [0180]
  • Expression vectors used in eukaryotic host cells (yeast, fungi, insect, plant, animal, human, or nucleated cells from other multicellular organisms) will also contain sequences necessary for the termination of transcription and for stabilizing the mRNA. Such sequences are commonly available from the 5′ and, occasionally 3′ , untranslated regions of eukaryotic or viral DNAs or cDNAs. These regions contain nucleotide segments transcribed as polyadenylated fragments in the untranslated portion of the mRNA encoding the polypeptide variant. One useful transcription termination component is the bovine growth hormone polyadenylation region. See WO94/11026 and the expression vector disclosed therein. [0181]
  • (vii) Selection and Transformation of Host Cells [0182]
  • Suitable host cells for cloning or expressing the DNA in the vectors herein are the prokaryote, yeast, or higher eukaryote cells described above. Suitable prokaryotes for this purpose include eubacteria, such as Gram-negative or Gram-positive organisms, for example, Enterobacteriaceae such as Escherichia, e.g., [0183] E coli, Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium, Serratia, e.g., Serratia marcescans, and Shigella, as well as Bacilli such as B. subtilis and B. licheniformis (e.g., B. licheniformis 41 P disclosed in DD 266,710 published Apr. 12, 1989), Pseudomonas such as P. aeruginosa, and Streptomyces. One preferred E. coil cloning host is E. coli 294 (ATCC 31,446), although other strains such as E. coli B, E. coil X1776 (ATCC 31,537), and E. coli W3110 (ATCC 27,325) are suitable. These examples are illustrative rather than limiting.
  • In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for polypeptide variant-encoding vectors. [0184] Saccharomyces cerevisiae, or common baker's yeast, is the most commonly used among lower eukaryotic host microorganisms. However, a number of other genera, species, and strains are commonly available and useful herein, such as Schizosaccharomyces pombe; Kluyveromyces hosts such as, e.g., K lactis, K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K. waltii (ATCC 56,500), K. drosophilarum (ATCC 36,906), K. thermotolerans, and K. marxianus; yarrowia (EP 402,226); Pichia pastoris (EP 183,070); Candida; Trichoderma reesia (EP 244,234); Neurospora crassa; Schwanniomyces such as Schwanniomyces occidentalis; and filamentous fungi such as, e.g., Neurospora, Penicillium, Tolypocladium, and Aspergillus hosts such as A. nidulans and A. niger.
  • Suitable host cells for the expression of glycosylated polypeptide variant are derived from multicellular organisms. Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains and variants and corresponding permissive insect host cells from hosts such as [0185] Spodoptera frugiperda (caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruiffly), and Bombyx mori have been identified. A variety of viral strains for transfection are publicly available, e.g., the L-1 variant of Autographa californica NPV and the Bm-5 strain of Bombyx mori NPV, and such viruses may be used as the virus herein according to the present invention, particularly for transfection of Spodoptera frugiperda cells.
  • Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato, and tobacco can also be utilized as hosts. [0186]
  • However, interest has been greatest in vertebrate cells, and propagation of vertebrate cells in culture (tissue culture) has become a routine procedure. Examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., [0187] J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells/-DHFR (CHO, Urlaub et al, Proc. Natl. Acad. Sci. USA 77:4216 (1980)); mouse sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; and a human hepatoma line (Hep G2).
  • Host cells are transformed with the above-described expression or cloning vectors for polypeptide variant production and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences. [0188]
  • (viii) Culturing the Host Cells [0189]
  • The host cells used to produce the polypeptide variant of this invention may be cultured in a variety of media. Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPMI-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing the host cells. In addition, any of the media described in Ham et al, [0190] Meth. Enz. 58:44 (1979), Barnes et al., Anal Biochem. 102:255 (1980), U.S. Pat. Nos. 4,767,704; 4,657,866; 4,927,762; 4,560,655; or 5,122,469; WO 90/03430; WO 87/00195; or U.S. Pat. No. Re. 30,985 may be used as culture media for the host cells. Any of these media may be supplemented as necessary with hormones and/or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics (such as GENTAMYCIN™ drug), trace elements (defined as inorganic compounds usually present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements may also be included at appropriate concentrations that would be known to those skilled in the art. The culture conditions, such as temperature, pH, and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.
  • (ix) Polypeptide Variant Purification [0191]
  • When using recombinant techniques, the polypeptide variant can be produced intracellularly, in the periplasmic space, or directly secreted into the medium. If the polypeptide variant is produced intracellularly, as a first step, the particulate debris, either host cells or lysed fragments, is removed, for example, by centrifugation or ultrafiltration. Carter et al., [0192] Bio/lTechnology 10:163-167 (1992) describe a procedure for isolating antibodies which are secreted to the periplasmic space of E coil. Briefly, cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30 min. Cell debris can be removed by centrifugation. Where the polypeptide variant is secreted into the medium, supernatants from such expression systems are generally first concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit. A protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants.
  • The polypeptide variant composition prepared from the cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being the preferred purification technique. The suitability of protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc region that is present in the polypeptide variant. Protein A can be used to purify polypeptide variants that are based on human γ1, γ2, or γ4 heavy chains (Lindmark et al, [0193] J. Immunol. Meth. 62:1-13 (1983)).
  • Protein G is recommended for all mouse isotypes and for human γ3 (Guss et al, [0194] EMBO J. 5:15671575 (1986)). The matrix to which the affinity ligand is attached is most often agarose, but other matrices are available. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose. Where the polypeptide variant comprises a C H3 domain, the Bakerbond ABX™ resin (J. T. Baker, Phillipsburg, N.J.) is useful for purification. Other techniques for protein purification such as fractionation on an ion-exchange column, ethanol precipitation, Reverse Phase HPLC, chromatography on silica, chromatography on heparin SEPHAROSE™ chromatography on an anion or cation exchange resin (such as a polyaspartic acid column), chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation are also available depending on the polypeptide variant to be recovered.
  • Following any preliminary purification step(s), the mixture comprising the polypeptide variant of interest and contaminants may be subjected to low pH hydrophobic interaction chromatography using an elution buffer at a pH between about 2.5-4.5, preferably performed at low salt concentrations (e.g.,from about 0-0.25M salt). [0195]
  • E. Pharmaceutical Formulations [0196]
  • Therapeutic formulations of the polypeptide variant are prepared for storage by mixing the polypeptide variant having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers ([0197] Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyidimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).
  • The formulation herein may also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. Such molecules are suitably present in combination in amounts that are effective for the purpose intended. [0198]
  • The active ingredients may also be entrapped in microcapsule prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsule and poly-(methylmethacylate) microcapsule, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). [0199]
  • The formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes. [0200]
  • Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the polypeptide variant, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and γ ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT™ (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(−)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods. When encapsulated antibodies remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37°C., resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S-S bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulihydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions. [0201]
  • F. Non-Therapeutic Uses for the Polypeptide Variant [0202]
  • The polypeptide variant of the invention may be used as an affinity purification agent. In this process, the polypeptide variant is immobilized on a solid phase such a Sephadex resin or filter paper, using methods well known in the art. The immobilized polypeptide variant is contacted with a sample containing the antigen to be purified, and thereafter the support is washed with a suitable solvent that will remove substantially all the material in the sample except the antigen to be purified, which is bound to the immobilized polypeptide variant. Finally, the support is washed with another suitable solvent, such as glycine buffer, pH 5.0, that will release the antigen from the polypeptide variant. [0203]
  • The polypeptide variant may also be useful in diagnostic assays, e.g., for detecting expression of an antigen of interest in specific cells, tissues, or serum. [0204]
  • For diagnostic applications, the polypeptide variant typically will be labeled with a detectable moiety. Numerous labels are available which can be generally grouped into the following categories: [0205]
  • (a) Radioisotopes, such as [0206] 35S, 14C, 125I, 3H, and 131I. The polypeptide variant can be labeled with the radioisotope using the techniques described in Current Protocots in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley-lnterscience, New York, New York, Pubs. (1991) for example and radioactivity can be measured using scintillation counting.
  • (b) Fluorescent labels such as rare earth chelates (europium chelates) or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin and Texas Red are available. The fluorescent labels can be conjugated to the polypeptide variant using the techniques disclosed in Current Protocols in Immunology, supra, for example. Fluorescence can be quantified using a fluorimeter. [0207]
  • (c) Various enzyme-substrate labels are available and U.S. Pat. No. 4,275,149 provides a review of some of these. The enzyme generally catalyzes a chemical alteration of the chromogenic substrate that can be measured using various techniques. For example, the enzyme may catalyze a color change in a substrate, which can be measured spectrophotometrically. Alternatively, the enzyme may alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying a change in fluorescence are described above. The chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light which can be measured (using a chemiluminometer, for example) or donates energy to a fluorescent acceptor. Examples of enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase; U.S. Pat. No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, β-galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like. Techniques for conjugating enzymes to antibodies are described in O′Sullivan et al., Methods for the Preparation of Enzyme-Antibody Conjugates for use in Enzyme Immunoassay, in Methods in Enzym. (ed J. Langone & H. Van Vunakis), Academic press, New York, 73:147-166 (1981). [0208]
  • Examples of enzyme-substrate combinations include, for example: [0209]
  • (i) Horseradish peroxidase (HRPO) with hydrogen peroxidase as a substrate, wherein the hydrogen peroxidase oxidizes a dye precursor (e.g.,orthophenylene diamine (OPD) or 3,3′,5,5′-tetramethyl benzidine hydrochloride (TMB)); [0210]
  • (ii) alkaline phosphatase (AP) with para-Nitrophenyl phosphate as chromogenic substrate; and [0211]
  • (iii) β-D-galactosidase (β-D-Gal) with a chromogenic substrate (e.g., p-nitrophenyl-β-D-galactosidase) or fluorogenic substrate 4-methylumbelliferyl-β-D-galactosidase. [0212]
  • Numerous other enzyme-substrate combinations are available to those skilled in the art. For a general review of these, see U.S. Pat. Nos. 4,275,149 and 4,318,980. [0213]
  • Sometimes, the label is indirectly conjugated with the polypeptide variant. The skilled artisan will be aware of various techniques for achieving this. For example, the polypeptide variant can be conjugated with biotin and any of the three broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the polypeptide variant in this indirect manner. Alternatively, to achieve indirect conjugation of the label with the polypeptide variant, the polypeptide variant is conjugated with a small hapten (e.g., digoxin) and one of the different types of labels mentioned above is conjugated with an anti-hapten polypeptide variant (e.g., anti-digoxin antibody). Thus, indirect conjugation of the label with the polypeptide variant can be achieved. [0214]
  • In another embodiment of the invention, the polypeptide variant need not be labeled, and the presence thereof can be detected using a labeled antibody which binds to the polypeptide variant. [0215]
  • The polypeptide variant of the present invention may be employed in any known assay method, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, [0216] Monoclonal Antibodies:A Manualof Techniques, pp.147-158 (CRC Press, Inc. 1987).
  • The polypeptide variant may also be used for in vivo diagnostic assays. Generally, the polypeptide variant is labeled with a radionuclide (such as [0217] 111In, 99Tc, 14C, 131I, 125I, 3H, 32 P or 35S) so that the antigen or cells expressing it can be localized using immunoscintiography.
  • G. In Vivo Uses for the Polypeptide Variant [0218]
  • It is contemplated that the polypeptide variant of the present invention may be used to treat a mammal e.g. a patient suffering from a disease or disorder who could benefit from administration of the polypeptide variant. The conditions which can be treated with the polypeptide variant are many and include cancer (e.g. where the polypeptide variant binds the HER2 receptor or CD20); allergic conditions such as asthma (with an anti-IgE antibody); and LFA-mediated disorders (e.g. where the polypeptide variant is an anti-LFA-1 or anti-ICAM-1 antibody) etc. Where the polypeptide variant does not bind complement, but retains FcR binding capability, exemplary diseases or disorders to be treated include: cancer (e.g. where ADCC function is desirable, but complement activation would lead to amplified side effects, such as vasculitis in the blood vessels at the tumor site); disorders treated with an agonist antibody; disorders wherein the polypeptide variant binds a soluble antigen and wherein stoichiometry leads to immune complexes which activate the complement cascade and result in unwanted side effects; conditions employing an antagonist antibody which downmodulates receptor function without damaging tissue or organ function; [0219]
  • intravenous immunoglobulin treatment for, e.g., immunodeficient individuals with autoimmune disorders. [0220]
  • The polypeptide variant is administered by any suitable means, including parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal, and, if desired for local immunosuppressive treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In addition, the polypeptide variant is suitably administered by pulse infusion, particularly with declining doses of the polypeptide variant. Preferably the dosing is given by injections, most preferably intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic. [0221]
  • For the prevention or treatment of disease, the appropriate dosage of polypeptide variant will depend on the type of disease to be treated, the severity and course of the disease, whether the polypeptide variant is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the polypeptide variant, and the discretion of the attending physician. The polypeptide variant is suitably administered to the patient at one time or over a series of treatments. [0222]
  • Depending on the type and severity of the disease, about 1 μg/kg to 15 mg/kg (e.g., 0.1-20 mg/kg) of polypeptide variant is an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion. A typical daily dosage might range from about 1 μg/kg to 100 mg/kg or more, depending on the factors mentioned above. For repeated administrations over several days or longer, depending on the condition, the treatment is sustained until a desired suppression of disease symptoms occurs. However, other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays. [0223]
  • The polypeptide variant composition will be formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The “therapeutically effective amount” of the polypeptide variant to be administered will be governed by such considerations, and is the minimum amount necessary to prevent, ameliorate, or treat a disease or disorder. The polypeptide variant need not be, but is optionally formulated with one or more agents currently used to prevent or treat the disorder in question. The effective amount of such other agents depends on the amount of polypeptide variant present in the formulation, the type of disorder or treatment, and other factors discussed above. These are generally used in the same dosages and with administration routes as used hereinbefore or about from 1 to 99% of the heretofore employed dosages. [0224]
  • The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of this invention. All literature and patent citations mentioned herein are expressly incorporated by reference. [0225]
    • EXAMPLE 1 Low Affinity Receptor Binding Assay
  • This assay determines binding of an IgG Fc region to recombinant FcγRIIa, FcγRIIb and FcγRIIIa α subunits expressed as His6-glutathione S transferase (GST)-tagged fusion proteins. Since the affinity of the Fc region of IgG1 for the FcγRI is in the nanomolar range, the binding of IgG1 Fc mutants can be measured by titrating monomeric IgG and measuring bound IgG with a polyclonal anti-lgG in a standard ELISA format (Example 2 below). The affinity of the other members of the FcγR family, ie. FcγRIIa, FcγRIIb and FcγRIIIa for IgG is however in the micromolar range and binding of monomeric IgG1 for these receptors can not be reliably measured in an ELISA format. [0226]
  • The following assay utilizes Fc mutants of recombinant anti-IgE E27 (FIGS. 4A and 4B) which, when mixed with human IgE at a 1:1 molar ratio, forms a stable hexamer consisting of three anti-IgE molecules and three IgE molecules. A recombinant chimeric form of IgE (chimeric IgE) was engineered and consists of a human IgE Fc region and the Fab of an anti-VEGF antibody (Presta et al [0227] Cancer Research 57:4593-4599 (1997)) which binds two VEGF molecules per mole of anti-VEGF. When recombinant human VEGF is added at a 2:1 molar ratio to chimeric IgE:E27 hexamers, the hexamers are linked into larger molecular weight complexes via the chimeric IgE Fab:VEGF interaction. The E27 component of this complex binds to the FcγRIIa, FcγRIIb and FcγIIIa α subunits with higher avidity to permit detection in an ELISA format.
    • MATERIALS AND METHODS
  • Receptor Coat. Fcγ receptor α subunits were expressed as GST fusions of His6 tagged extracellular domains (ECDs) in 293 cells resulting in an ECD-6His-GST fusion protein (Graham et al [0228] J. Gen. Virol. 36:59-72 (1977) and Gorman et al DNA Prot. Eng. Tech. 2:3-10 (1990)) and purified by Ni-NTA column chromatography (Qiagen, Australia) and buffer exchanged into phosphate buffered saline (PBS). Concentrations were determined by absorption at 280nm using extinction coefficients derived by amino acid composition analysis. Receptors were coated onto Nunc F96 maxisorb plates (cat no. 439454) at 100 ng per well by adding 100 μl of receptor-GST fusion at 1 μg/ml in PBS and incubated for 48 hours at 4° C. Prior to assay, plates are washed 3×0 with 250 μl of wash buffer (PBS pH 7.4 containing 0.5% TWEEN 20™) and blocked with 250 μl of assay buffer (50 mM Tris buffered saline, 0.05% TWEEN 20™, 0.5% RIA grade bovine albumin (Sigma A7888), and 2 mM EDTA pH 7.4).
  • Immune Complex Formation: Equal molar amounts (1:1) of E27 and recombinant chimeric IgE which binds two moles recombinant human VEGF per mole of chimeric IgE are added to a 12 ×75 mm polypropylene tube in PBS and mixed by rotation for 30 minutes at 25° C. E27 (anti-IgE) /chimeric IgE (IgE) hexamers are formed during this incubation. Recombinant human VEGF (165 form, MW 44,000) is added at a 2:1 molar ratio to the IgE concentration and mixed by rotation an additional 30 minutes at 25° C. VEGF- chimeric IgE binding links E27:chimeric IgE hexamers into larger molecular weight complexes which bind FcγR α subunit ECD coated plates via the Fc region of the E27 antibody. [0229]
  • E27:chimeric IgE:VEGF(1:1:2 molar ratio) complexes are added to FcγR α subunit coated plates at E27 concentrations of 5 μg and 1 μg total IgG in quadruplicate in assay buffer and incubated for 120 minutes at 25° C. on an orbital shaker. [0230]
  • Complex Detection: Plates are washed 5× with wash buffer to remove unbound complexes and IgG binding is detected by adding 100 μl horse radish peroxidase (HRP) conjugated goat anti-human IgG (γ) heavy chain specific (Boehringer Mannheim 1814249) at 1:10,000 in assay buffer and incubated for 90 min at 25° C. on an orbital shaker. Plates are washed 5× with wash buffer to remove unbound HRP goat anti-human IgG and bound anti-lgG is detected by adding 100 μl of substrate solution (0.4mg/ml o-phenylenedaimine dihydrochloride, Sigma P6912, 6 mM H[0231] 2O2 in PBS) and incubating for 8 min at 25° C. Enzymatic reaction is stopped by the addition of 100 μl 4.5N H2SO4 and colorimetric product is measured at 490 nm on a 96 well plate densitometer (Molecular Devices). Binding of E27 mutant complexes is expressed as a percent of the wild type E27 containing complex.
    • EXAMPLE 2 Identification of Unique C1q Binding Sites in a Human IgG Antibody
  • In the present study, mutations were identified in the CH2 domain of a human IgG1 antibody, “C2B8” (Reff et al., Blood 83:435 (1994)), that ablated binding of the antibody to C1q but did not alter the conformation of the antibody nor affect binding to each of the FcγRs. By alanine scanning mutagenesis, three mutants in human IgG1 were identified, K322A P329A, and P331, that were non-lytic and had decreased binding to C1q. The data suggested that the core C1q binding sites in human IgG1 is different from that of murine IgG2b. In addition, K322A, P329A and P331A were found to bind normally to the CD20 antigen, and to four Fc receptors, FcγRI, FcγRII, FcγRIII and FcRn. [0232]
    • MATERIALS AND METHODS
  • Construction of C2B8 Mutants: The chimeric light and heavy chains of anti-CD20 antibody C2B8 (Reff et al., [0233] Blood 83:435 (1994)) subcloned separately into previously described PRK vectors (Gorman et al, DNA Protein Eng. Tech. 2:3 (1990)) were used. By site directed mutagenesis (Kunkel et al., Proc. Natl. Acad.Sci.USA 82:488 (1985)), alanine scan variants of the Fc region in the heavy chain were constructed. The heavy and light chain plasmids were co-transfected into an adenovirus transformed human embryonic kidney cell line as previously described (Werther et al., J. Immunol 157:4986 (1996)). The media was changed to serum-free 24 hours after transfection and the secreted antibody was harvested after five days. The antibodies were purified using Protein A-SEPHAROSE CL-4B™ (Pharmacia), buffer exchanged and concentrated to 0.5 ml with PBS using a Centricon-30 (Amicon), and stored at 4° C. The concentration of the antibody was determined using total Ig-binding ELISA.
  • C1q Binding ELISA: Costar 96 well plates were coated overnight at 4° C. with the indicated concentrations of 2B8 in coating buffer (0.05 M sodium carbonate buffer), pH 9. The plates were then washed 3× with PBS/ 0.05[0234] % TWEEN 20™, pH 7.4 and blocked with 200 μl of ELISA diluent without thimerosal (0.1 M NaPO4/0.1M NaCl/0.1% gelatin/0.05% TWEEN 20 ™/ 0.05% ProClin300) for 1 hr at room temperature. The plate was washed 3× with wash buffer, an aliquot of 100 μl of 2 μg/ml C1q (Quidel, San Diego, Calif.) was added to each well and incubated for 2 hrs at room temperature. The plate was then washed 6× with wash buffer. 100 μl of a 1:1000 dilution of sheep anti-complement C1q peroxidase conjugated antibody (Biodesign) was added to each well and incubated for 1 hour at room temperature. The plate was again washed 6× with wash buffer and 100 μl of substrate buffer (PBS/0.012% H2O2) containing OPD (O-phenylenediamine dihydrochloride (Sigma)) was added to each well. The oxidation reaction, observed by the appearance of a yellow color, was allowed to proceed for 30 minutes and stopped by the addition of 100 μl of 4.5 N H2SO4. The absorbance was then read at (492-405) nm using a microplate reader (SPECTRA MAX 250™, Molecular Devices Corp.). The appropriate controls were run in parallel (i.e. the ELISA was performed without C1q for each concentration of C2B8 used and also the ELISA was performed without C2B8). For each mutant, C1q binding was measured by plotting the absorbance (492-405) nm versus concentration of C2B8 in μg/ml using a 4-parameter curve fitting program (KALEIDAGRAPHTM) and comparing EC50 values.
  • Complement Dependent Cytotoxicity (CDC) Assay: This assay was performed essentially as previously described (Gazzano-Santoro et al, [0235] J. Immunol. Methods 202:163 (1997)). Various concentrations of C2B8 (0.08-20 μg/ml) were diluted with RHB buffer (RPMI 1640/20 mM HEPES (pH 7.2)/2mM Glutamine/0.1% BSA/100 μml Gentamicin). Human complement (Quidel) was diluted 1:3 in RHB buffer and WIL2-S cells (available from the ATCC, Manassas, Va.) which express the CD20 antigen were diluted to a density of 1×106 cells /ml with RHB buffer. Mixtures of 150 μl containing equal volumes of C2B8, diluted human complement and WIL2-S cells were added to a flat bottom tissue culture 96 well plate and allowed to incubate for 2 hrs at 37° C. and 5% CO2 to facilitate complement mediated cell lysis. 50 μl of alamar blue (Accumed International) was then added to each well and incubated overnight at 37° C. The absorbance was measured using a 96-well fluorometer with excitation at 530 nm and emission at 590 nm. As described by Gazzano-Santoro et al., the results are expressed in relative fluorescence units (RFU). The sample concentrations were computed from a C2B8 standard curve and the percent activity as compared to wild type C2B8 is reported for each mutant.
  • CD20 Binding Potency of the C2B8 Mutants: The binding of C2B8 and mutants to the CD20 antigen were assessed by a method previously described (Reff et al, (1994), supra; reviewed in Gazzano-Santoro et al, (1996), supra). WIL2-S cells were grown for 3-4 days to a cell density of 1×10[0236] 6 cells/ml. The cells were washed and spun twice in FACS buffer (PBS/0.1% BSA/0.02% NaN3) and resuspended to a cell density of 5×106 cells/ml. 200 μl of cells (5×106 cells/ml) and 20 μl of diluted C2B8 samples were added to a 5 ml tube and incubated at room temperature for 30 minutes with agitation. The mixture was then washed with 2 ml of cold FACS buffer, spun down and resuspended in 200 μl of cold FACS buffer. To the suspension, 10 μl of goat anti-human IgG-FITC (American Qualex Labs.) was added and the mixture was incubated in the dark at room temperature for 30 minutes with agitation. After incubation, the mixture was washed with 2 ml of FACS buffer, spun down and resuspended in 1 ml of cold fixative buffer (1% formaldehyde in PBS). The samples were analyzed by flow cytometry and the results expressed as relative fluorescence units (RFU) were plotted against antibody concentrations using a 4-parameter curve fitting program (KALEIDAGRAPH™). The EC50 values are reported as a percentage of that of the C2B8 reference material.
  • FcγR Binding Eilsas: [0237]
  • FcγRI α subunit-GST fusion was coated onto Nunc F96 maxisorb plates (cat no. 439454) by adding 100 μl of receptor-GST fusion at 1 μg/ml in PBS and incubated for 48 hours at 4° C. Prior to assay, plates are washed 3× with 250 μl of wash buffer (PBS pH 7.4 containing 0.5[0238] % TWEEN 20™) and blocked with 250 μl of assay buffer (50 mM Tris buffered saline, 0.05% TWEEN 20™, 0.5% RIA grade bovine albumin (Sigma A7888), and 2 mM EDTA pH 7.4). Samples diluted to 10 μg/ml in 1 ml of assay buffer are added to FcγRI α subunit coated plates and incubated for 120 minutes at 25° C. on an orbital shaker. Plates are washed 5× with wash buffer to remove unbound complexes and IgG binding is detected by adding 100 μl horse radish peroxidase (HRP) conjugated goat anti-human IgG (γ) heavy chain specific (Boehringer Mannheim 1814249) at 1:10,000 in assay buffer and incubated for 90 min at 25° C. on an orbital shaker. Plates are washed 5× with wash buffer to remove unbound HRP goat anti-human IgG and bound anti-lgG is detected by adding 100 μl of substrate solution (0.4mg/ml o-phenylenedaimine dihydrochloride, Sigma P6912, 6 mM H2O2 in PBS) and incubating for 8 min at 25° C. Enzymatic reaction is stopped by the addition of 100 μl 4.5N H2SO4 and colorimetric product is measured at 490 nm on a 96 well plate densitometer (Molecular Devices). Binding of variant is expressed as a percent of the wild type molecule.
  • FcγRII and III binding ELISAs were performed as described in Example 1 above. [0239]
  • For measuring FcRn binding activity of IgG variants, ELISA plates were coated with 2 μg/ml streptavidin (Zymed, South San Francisco) in 50 mM carbonate buffer, pH 9.6, at 4° C. overnight and blocked with PBS-0.5% BSA, pH 7.2 at room temperature for one hour. Biotinylated FcRn (prepared using biotin-X-NHS from Research Organics, Cleveland, Ohio and used at 1-2 μg/ml) in PBS-0.5% BSA, 0.05[0240] % polysorbate 20, pH 7.2, was added to the plate and incubated for one hour. Two fold serial dilutions of IgG standard (1.6-100 ng/ml) or variants in PBS-0.5% BSA, 0.05% polysorbate 20, pH 6.0, were added to the plate and incubated for two hours. Bound IgG was detected using peroxidase labeled goat F(ab′)2 anti-human lgG F(ab′)2 in the above pH 6.0 buffer (Jackson lmmunoResearch, West Grove, Pa.) followed by 3,3′,5,5′-tetramethyl benzidine (Kirgaard & Perry Laboratories) as the substrate. Plates were washed between steps with PBS-0.05% polysorbate 20 at either pH 7.2 or 6.0. Absorbance was read at 450 nm on a Vmax plate reader (Molecular Devices, Menlo Park, Calif.). Titration curves were fit with a four-parameter nonlinear regression curve-fitfing program (KaleidaGraph, Synergy software, Reading, Pa.). Concentrations of IgG variants corresponding to the mid-point absorbance of the titration curve of the standard were calculated and then divided by the concentration of the standard corresponding to the mid-point absorbance of the standard titration curve.
    • RESULTS AND DISCUSSION
  • By alanine scanning mutagenesis, several single point mutations were constructed in the CH2 domain of C2B8 beginning with E318A, K320A and K322A. All the mutants constructed bound normally to the CD20 antigen (Table 1). [0241]
    TABLE 1
    wt E318A K320A K322A P329A P331A
    FcRn + + + +
    CD20 + + + + + +
    FcγRI + + + + + +
    FcγRII + + + + + +
    FcγRIII + + + + + +
    *C1q +++ ++ +++ − − −
    CDC + + + − − −
  • Where binding of human complement to an antibody with a human Fc was analyzed, the ability of E318A and K320A to activate complement was essentially identical to that of wild type C2B8 (Table 1). When compared to wild type C2B8, there appears to be little difference in the binding of E318A and K320A to C1q. There is only a 10% decrease in the binding of K320A and about a 30% decrease in the binding of E318A to C1q (FIG. 2). The results indicate that the effect of the E318A and the K320A substitution on complement activation and C1q binding is minimal. Also, the human IgG1 of C2B8 was substituted for human IgG2 and used as a negative control in the C1q binding studies. The IgG2 mutant appears to have a much lower affinity for C1q than the E318A and K320A mutants (FIG. 2). Thus, the results demonstrate that E318 and K320 do not constitute the core C1q binding sites for human IgG1. Conversely, the K322A substitution had a significant effect on both complement activity and C1q binding. The K322A mutant did not have CDC activity when tested in the above CDC assay and was more than a 100 fold lower than wild type C2B8 in binding to C1q (FIG. 2). In the human system, K322 is the only residue of the proposed core C1q binding sites that appeared to have a significant effect on complement activation and C1q binding. [0242]
  • Since the Duncan and Winter study was performed using mouse IgG2b and the above results reveal that K320 and E318 in human IgG1 are not involved in C1q binding, and without being bound to any one theory, the above data suggest that the C1q binding region in murine IgGs is different from that of the human. To investigate this further and also to identify additional mutants that do not bind to C1 q and hence do not activate complement, several more point mutations in the vicinity of K322 were constructed as assessed from the three dimensional structure of the C2B8 Fc. Mutants constructed, K274A, N276A, Y278A, S324A, P329A, P331A, K334A, and T335A, were assessed for their ability to bind C1q and also to activate complement. Many of these substitutions had little or no effect on C1q binding or complement activation. In the above assays, the P329 A and the P331 A mutants did not activate complement and had decreased binding to C1q. The P331 A mutant did not activate complement and was 60 fold lower in binding to C1q (FIG. 3) when compared to wild type C2B8 (FIG. 2). The concentration range of the antibody variants used in FIG. 3 is expanded to 100 μg/ml in order to observe saturation of C1q binding to the P331 A variant. The mutation P329A results in an antibody that does not activate complement and is more than a 100 fold lower in binding to C1q (FIG. 3) when compared to wild type C2B8 (FIG. 2). [0243]
  • Mutants that did not bind to C1q and hence did not activate complement were examined for their ability to bind to the Fc receptors: FcγRI, FcγRIIa, FcγRIlb, FcγRIIIa and FcRn. This particular study was performed using a humanized anti-IgE antibody, an IgG1 antibody with these mutations (see Example 1 above). The results revealed the mutants, K322A and P329A, bind to all the Fc receptors to the same extent as the wild type protein (Table 2). However, there was a slight decrease in the binding of P331 A to FcγRIIb. [0244]
  • In conclusion, two amino acid substitutions in the COOH terminal region of the CH2 domain of human IgG1, K322A and P329A were identified that result in more than 100 fold decrease in C1q binding and do not activate the CDC pathway. These two mutants, K322A and P329A, bind to all Fc receptors with the same affinity as the wild type antibody. Based on the results, summarized in Table 2, and without being bound to any one theory, it is proposed that the C1q binding epicenter of human IgG1 is centered around K322, P329 and P331 and is different from the murine IgG2b epicenter which constitutes E318, K320 and K322. [0245]
    TABLE 2
    wt E318A K320A K322A P329A P331A
    CD20
    100 89 102 86 112 103 
    a FcγRI 100 93 102 90 104 74
    a FcγRIIa 100 113 94 109 111 86
    a FcγRIIb 100 106 83 101 96 58
    a FcγIII 100 104 72 90 85 73
    CDC 100 108 108 none none none
  • [0246]
  • 1 2 1 218 PRT Artificial Sequence Artificial Sequence 1-218 Sequence is completely synthesized 1 Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val 1 5 10 15 Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Lys Pro Val Asp 20 25 30 Gly Glu Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly 35 40 45 Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Tyr Leu Glu Ser 50 55 60 Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 65 70 75 Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 80 85 90 Tyr Cys Gln Gln Ser His Glu Asp Pro Tyr Thr Phe Gly Gln Gly 95 100 105 Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe 110 115 120 Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser 125 130 135 Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val 140 145 150 Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 155 160 165 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 170 175 180 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 185 190 195 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr 200 205 210 Lys Ser Phe Asn Arg Gly Glu Cys 215 218 2 451 PRT Artificial Sequence Artificial Sequence 1-451 Sequence is completely synthesized 2 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 1 5 10 15 Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Tyr Ser Ile Thr 20 25 30 Ser Gly Tyr Ser Trp Asn Trp Ile Arg Gln Ala Pro Gly Lys Gly 35 40 45 Leu Glu Trp Val Ala Ser Ile Lys Tyr Ser Gly Glu Thr Lys Tyr 50 55 60 Asn Pro Ser Val Lys Gly Arg Ile Thr Ile Ser Arg Asp Asp Ser 65 70 75 Lys Asn Thr Phe Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 80 85 90 Thr Ala Val Tyr Tyr Cys Ala Arg Gly Ser His Tyr Phe Gly His 95 100 105 Trp His Phe Ala Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser 110 115 120 Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser 125 130 135 Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 140 145 150 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 155 160 165 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 170 175 180 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 185 190 195 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 200 205 210 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 215 220 225 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 275 280 285 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 290 295 300 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 305 310 315 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 320 325 330 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 335 340 345 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu 350 355 360 Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 365 370 375 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 380 385 390 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 395 400 405 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 410 415 420 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 425 430 435 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 440 445 450 Lys 451

Claims (23)

1. A variant of a polypeptide comprising a human IgG Fc region, which variant comprises an amino acid substitution at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
2. The variant of claim 1 having Pro329 of the human IgG Fc region substituted with another amino acid.
3. The variant of claim 1 wherein the polypeptide comprises an antibody.
4. The variant of claim 1 wherein the polypeptide comprises a human IgG1 Fc region.
5. The variant of claim 1 which does not activate complement.
6. The variant of claim 5 which binds an FcR.
7. The variant of claim 6 which binds FcγRI, FcγRII, FcγRIII and FcRn.
8. A variant of a polypeptide comprising a human IgG Fc region, which variant binds FcγRI, FcγRII, FcγRIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 322 or amino acid position 329, or both amino acid positions of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
9. A composition comprising the variant of claim 1 and a physiologically acceptable carrier.
10. A method for modifying a polypeptide comprising a human IgG Fc region comprising substituting an amino acid residue at amino acid position 329, or at two or all of amino acid positions 329, 331 and 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
11. Isolated nucleic acid encoding the variant of claim 1.
12. A vector comprising the nucleic acid of claim 11.
13. A host cell comprising the vector of claim 12.
14. A process of producing an polypeptide variant comprising culturing the host cell of claim 13 so that the nucleic acid is expressed.
15. The process of claim 14 further comprising recovering the polypeptide variant from the host cell culture.
16. A method of treating a mammal suffering from a disorder comprising administering to the mammal a therapeutically effective amount of a variant of a polypeptide comprising a human IgG Fc region, which variant binds FcγRI, FcγRII, FcγRIII and FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 329, 331 or 322 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat.
17. An immune complex comprising:
(a) an Fc region-containing polypeptide;
(b) a first target molecule which comprises at least two binding sites for the Fc region-containing polypeptide; and
(c) a second target molecule which comprises at least two binding sites for the first target molecule.
18. A method for determining binding of an analyte to a receptor comprising the following steps performed sequentially:
(a) forming a molecular complex between the analyte and a first target molecule, wherein the first target molecule comprises at least two binding sites for the analyte; and
(b) determining binding of the molecular complex of step (a) to the receptor.
19. The method of claim 18 wherein the analyte is an Fc region-containing polypeptide.
20. The method of claim 18 wherein the receptor is a low affinity receptor.
21. The method of claim 18 wherein the receptor is an FcR.
22. The method of claim 18 wherein the molecular complex of step (a) further comprises a second target molecule which comprises at least two binding sites for the first target molecule.
23. A kit useful for determining binding of an analyte to a receptor comprising:
(a) a first target molecule which comprises at least two binding sites for the analyte; and
(b) a second target molecule which comprises at least two binding sites for the first target molecule.
US09/792,938 1998-04-02 2001-02-26 Polypeptide variants Abandoned US20030166868A1 (en)

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Cited By (55)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030158389A1 (en) * 1998-04-02 2003-08-21 Genentech, Inc. Polypeptide variants
US20040132101A1 (en) * 2002-09-27 2004-07-08 Xencor Optimized Fc variants and methods for their generation
US20040191244A1 (en) * 1999-01-15 2004-09-30 Genentech, Inc. Polypeptide variants with altered effector function
US20050069521A1 (en) * 2003-08-28 2005-03-31 Emd Lexigen Research Center Corp. Enhancing the circulating half-life of interleukin-2 proteins
US20050164352A1 (en) * 2003-12-30 2005-07-28 Emd Lexigen Research Center Corp. IL-7 fusion proteins
WO2005070967A2 (en) * 2004-01-22 2005-08-04 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US20050244403A1 (en) * 2004-03-24 2005-11-03 Xencor, Inc. Immunoglobulin variants outside the Fc region
US20050249723A1 (en) * 2003-12-22 2005-11-10 Xencor, Inc. Fc polypeptides with novel Fc ligand binding sites
US20060074225A1 (en) * 2004-09-14 2006-04-06 Xencor, Inc. Monomeric immunoglobulin Fc domains
US20060173170A1 (en) * 2004-11-12 2006-08-03 Xencor, Inc. Fc variants with altered binding to FcRn
US20060194291A1 (en) * 1999-01-15 2006-08-31 Genentech, Inc. Polypeptide variants with altered effector function
US20060235208A1 (en) * 2002-09-27 2006-10-19 Xencor, Inc. Fc variants with optimized properties
US20060275282A1 (en) * 2005-01-12 2006-12-07 Xencor, Inc. Antibodies and Fc fusion proteins with altered immunogenicity
US20060275283A1 (en) * 2003-11-12 2006-12-07 Biogen Idec Ma Inc. Fcgamma receptor-binding polypeptide variants and methods related thereto
US20070003546A1 (en) * 2002-03-01 2007-01-04 Xencor, Inc. Optimized Fc variants and methods for their generation
US7183387B1 (en) 1999-01-15 2007-02-27 Genentech, Inc. Polypeptide variants with altered effector function
US20070148171A1 (en) * 2002-09-27 2007-06-28 Xencor, Inc. Optimized anti-CD30 antibodies
US20070219133A1 (en) * 2002-03-01 2007-09-20 Xencor, Inc. CD52 OPTIMIZED Fc VARIANTS AND METHODS FOR THEIR GENERATION
US20070238665A1 (en) * 2003-03-03 2007-10-11 Xencor, Inc. Fc Variants Having Decreased Affinity for FcyRIIc
US20070275460A1 (en) * 2003-03-03 2007-11-29 Xencor.Inc. Fc Variants With Optimized Fc Receptor Binding Properties
US20080057056A1 (en) * 2003-03-03 2008-03-06 Xencor, Inc. Fc Variants with Increased Affinity for FcyRIIC
US20080206867A1 (en) * 2005-10-03 2008-08-28 Desjarlais John R Fc variants with optimized Fc receptor binding properties
US20080254027A1 (en) * 2002-03-01 2008-10-16 Bernett Matthew J Optimized CD5 antibodies and methods of using the same
US20080260731A1 (en) * 2002-03-01 2008-10-23 Bernett Matthew J Optimized antibodies that target cd19
US20080267976A1 (en) * 2005-10-06 2008-10-30 Gregory Alan Lazar Optimized Anti-Cd30 Antibodies
US20080313379A1 (en) * 2007-06-15 2008-12-18 United Memories, Inc. Multiple bus charge sharing
US20090136936A1 (en) * 2007-05-01 2009-05-28 George Georgiou Immunoglobulin fc libraries
US20100104557A1 (en) * 2006-09-18 2010-04-29 Xencor, Inc. Optimized Antibodies that Target HM1.24
US7820161B1 (en) 1999-05-07 2010-10-26 Biogen Idec, Inc. Treatment of autoimmune diseases
US20100272723A1 (en) * 2006-08-14 2010-10-28 Xencor, Inc. Optimized Antibodies that Target CD19
US20100330076A1 (en) * 2009-06-30 2010-12-30 George Georgiou Immunoglobulin fc polypeptides
US8084582B2 (en) 2003-03-03 2011-12-27 Xencor, Inc. Optimized anti-CD20 monoclonal antibodies having Fc variants
US8101720B2 (en) 2004-10-21 2012-01-24 Xencor, Inc. Immunoglobulin insertions, deletions and substitutions
US8188231B2 (en) 2002-09-27 2012-05-29 Xencor, Inc. Optimized FC variants
US8318907B2 (en) 2004-11-12 2012-11-27 Xencor, Inc. Fc variants with altered binding to FcRn
US8409577B2 (en) 2006-06-12 2013-04-02 Emergent Product Development Seattle, Llc Single chain multivalent binding proteins with effector function
US8546543B2 (en) 2004-11-12 2013-10-01 Xencor, Inc. Fc variants that extend antibody half-life
US8802820B2 (en) 2004-11-12 2014-08-12 Xencor, Inc. Fc variants with altered binding to FcRn
US8853366B2 (en) 2001-01-17 2014-10-07 Emergent Product Development Seattle, Llc Binding domain-immunoglobulin fusion proteins
US8969526B2 (en) 2011-03-29 2015-03-03 Roche Glycart Ag Antibody Fc variants
US9051373B2 (en) 2003-05-02 2015-06-09 Xencor, Inc. Optimized Fc variants
US9101609B2 (en) 2008-04-11 2015-08-11 Emergent Product Development Seattle, Llc CD37 immunotherapeutic and combination with bifunctional chemotherapeutic thereof
US9238878B2 (en) 2009-02-17 2016-01-19 Redwood Bioscience, Inc. Aldehyde-tagged protein-based drug carriers and methods of use
US9475881B2 (en) 2010-01-19 2016-10-25 Xencor, Inc. Antibody variants with enhanced complement activity
US9540438B2 (en) 2011-01-14 2017-01-10 Redwood Bioscience, Inc. Aldehyde-tagged immunoglobulin polypeptides and methods of use thereof
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US9714282B2 (en) 2003-09-26 2017-07-25 Xencor, Inc. Optimized Fc variants and methods for their generation
US10143748B2 (en) 2005-07-25 2018-12-04 Aptevo Research And Development Llc B-cell reduction using CD37-specific and CD20-specific binding molecules
US10457737B2 (en) 2015-02-09 2019-10-29 Research Development Foundation Engineered immunoglobulin Fc polypeptides displaying improved complement activation
US10654916B2 (en) 2011-04-21 2020-05-19 The Regents Of The University Of California, A California Corporation Compositions and methods for the treatment of neuromyelitis optica
US11208632B2 (en) 2016-04-26 2021-12-28 R.P. Scherer Technologies, Llc Antibody conjugates and methods of making and using the same
US11352426B2 (en) 2015-09-21 2022-06-07 Aptevo Research And Development Llc CD3 binding polypeptides
US11401348B2 (en) 2009-09-02 2022-08-02 Xencor, Inc. Heterodimeric Fc variants
US11820830B2 (en) 2004-07-20 2023-11-21 Xencor, Inc. Optimized Fc variants
US11932685B2 (en) 2007-10-31 2024-03-19 Xencor, Inc. Fc variants with altered binding to FcRn

Families Citing this family (1482)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080050367A1 (en) * 1998-04-07 2008-02-28 Guriq Basi Humanized antibodies that recognize beta amyloid peptide
US6761888B1 (en) * 2000-05-26 2004-07-13 Neuralab Limited Passive immunization treatment of Alzheimer's disease
TWI239847B (en) * 1997-12-02 2005-09-21 Elan Pharm Inc N-terminal fragment of Abeta peptide and an adjuvant for preventing and treating amyloidogenic disease
US7964192B1 (en) 1997-12-02 2011-06-21 Janssen Alzheimer Immunotherapy Prevention and treatment of amyloidgenic disease
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US6194551B1 (en) * 1998-04-02 2001-02-27 Genentech, Inc. Polypeptide variants
GB9809951D0 (en) 1998-05-08 1998-07-08 Univ Cambridge Tech Binding molecules
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DE60142614D1 (en) * 2000-01-27 2010-09-02 Medimmune Inc INITÄT
CA2396793A1 (en) * 2000-02-16 2001-08-23 Genentech, Inc. Uses of agonists and antagonists to modulate activity of tnf-related molecules
DK1259547T3 (en) * 2000-03-01 2012-10-15 Medimmune Inc HIGH POTENTIAL, RECOMBINANT ANTIBODIES AND PROCEDURE FOR PRODUCING THEM
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US6787524B2 (en) * 2000-09-22 2004-09-07 Tanox, Inc. CpG oligonucleotides and related compounds for enhancing ADCC induced by anti-IgE antibodies
US7179900B2 (en) * 2000-11-28 2007-02-20 Medimmune, Inc. Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment
US6855493B2 (en) 2000-11-28 2005-02-15 Medimmune, Inc. Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment
US7700751B2 (en) 2000-12-06 2010-04-20 Janssen Alzheimer Immunotherapy Humanized antibodies that recognize β-amyloid peptide
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EP2341060B1 (en) * 2000-12-12 2019-02-20 MedImmune, LLC Molecules with extended half-lives, compositions and uses thereof
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US7829084B2 (en) * 2001-01-17 2010-11-09 Trubion Pharmaceuticals, Inc. Binding constructs and methods for use thereof
US20030133939A1 (en) * 2001-01-17 2003-07-17 Genecraft, Inc. Binding domain-immunoglobulin fusion proteins
US20040058445A1 (en) * 2001-04-26 2004-03-25 Ledbetter Jeffrey Alan Activation of tumor-reactive lymphocytes via antibodies or genes recognizing CD3 or 4-1BB
CN1636067A (en) * 2001-08-03 2005-07-06 杰南技术公司 TACIs and BR3 polypeptides and uses thereof
US20060073095A1 (en) * 2001-08-22 2006-04-06 Steven Kessler Methods for screening antibody-producing cells on heterogeneous antigen substrates
US20040235068A1 (en) * 2001-09-05 2004-11-25 Levinson Arthur D. Methods for the identification of polypeptide antigens associated with disorders involving aberrant cell proliferation and compositions useful for the treatment of such disorders
US20110045005A1 (en) 2001-10-19 2011-02-24 Craig Crowley Compositions and methods for the treatment of tumor of hematopoietic origin
WO2003035835A2 (en) * 2001-10-25 2003-05-01 Genentech, Inc. Glycoprotein compositions
US6911321B2 (en) * 2001-12-19 2005-06-28 Genentech, Inc. Non-human primate Fc receptors and methods of use
AU2003216250A1 (en) * 2002-02-11 2003-09-04 Genentech, Inc. Antibody variants with faster antigen association rates
US20040002587A1 (en) * 2002-02-20 2004-01-01 Watkins Jeffry D. Fc region variants
US20090042291A1 (en) * 2002-03-01 2009-02-12 Xencor, Inc. Optimized Fc variants
US20100311954A1 (en) * 2002-03-01 2010-12-09 Xencor, Inc. Optimized Proteins that Target Ep-CAM
MY139983A (en) * 2002-03-12 2009-11-30 Janssen Alzheimer Immunotherap Humanized antibodies that recognize beta amyloid peptide
US20030219436A1 (en) * 2002-03-15 2003-11-27 Ledbetter Jeffrey A. Compositions and methods to regulate an immune response using CD83 gene expressed in tumors and using soluble CD83-Ig fusion protein
DE60334453D1 (en) * 2002-05-30 2010-11-18 Macrogenics Inc CD16A BINDING PROTEINS AND USE FOR THE TREATMENT OF IMMUNE DISEASES
US7132100B2 (en) * 2002-06-14 2006-11-07 Medimmune, Inc. Stabilized liquid anti-RSV antibody formulations
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US8946387B2 (en) * 2002-08-14 2015-02-03 Macrogenics, Inc. FcγRIIB specific antibodies and methods of use thereof
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US8968730B2 (en) 2002-08-14 2015-03-03 Macrogenics Inc. FcγRIIB specific antibodies and methods of use thereof
US8187593B2 (en) * 2002-08-14 2012-05-29 Macrogenics, Inc. FcγRIIB specific antibodies and methods of use thereof
BRPI0314814C1 (en) * 2002-09-27 2021-07-27 Xencor Inc antibody comprising an fc variant
NZ568769A (en) 2002-10-17 2010-04-30 Genmab As Human monoclonal antibodies against CD20
DE10254601A1 (en) 2002-11-22 2004-06-03 Ganymed Pharmaceuticals Ag Gene products differentially expressed in tumors and their use
AR042485A1 (en) * 2002-12-16 2005-06-22 Genentech Inc HUMANIZED ANTIBODY THAT JOINS THE HUMAN CD20
EP2368578A1 (en) 2003-01-09 2011-09-28 Macrogenics, Inc. Identification and engineering of antibodies with variant Fc regions and methods of using same
US7960512B2 (en) * 2003-01-09 2011-06-14 Macrogenics, Inc. Identification and engineering of antibodies with variant Fc regions and methods of using same
EP2272533A1 (en) 2003-01-13 2011-01-12 MacroGenics, Inc. Soluble FcyR fusion proteins and methods of use thereof
UA87453C2 (en) * 2003-02-01 2009-07-27 Вайет Use a fragment ав for treatment of alzheimer's disease
CN1832965B (en) 2003-03-04 2011-03-02 麒麟麦酒株式会社 Endothelial cell specific antibodies and uses thereof
UA99933C2 (en) 2003-04-09 2012-10-25 Дженентек, Инк. Therapy of autoimmune disease in a patient with an inadequate response to tnf-alpha inhibitor
TWI353991B (en) 2003-05-06 2011-12-11 Syntonix Pharmaceuticals Inc Immunoglobulin chimeric monomer-dimer hybrids
CA2897608C (en) * 2003-05-09 2018-07-31 Duke University Cd20-specific antibodies and methods employing same
TWI374893B (en) * 2003-05-30 2012-10-21 Janssen Alzheimer Immunotherap Humanized antibodies that recognize beta amyloid peptide
US20050163775A1 (en) * 2003-06-05 2005-07-28 Genentech, Inc. Combination therapy for B cell disorders
CA2526402A1 (en) * 2003-06-05 2005-01-20 Genentech, Inc. Blys antagonists and uses thereof
US20040254108A1 (en) * 2003-06-13 2004-12-16 Jing Ma Preparation and application of anti-tumor bifunctional fusion proteins
US20050232931A1 (en) * 2003-06-13 2005-10-20 Oncomax Acquisition Corp. Preparation and application of anti-tumor bifunctional fusion proteins
WO2005003175A2 (en) * 2003-06-13 2005-01-13 Biogen Idec Ma Inc. Aglycosyl anti-cd154 (cd40 ligand) antibodies and uses thereof
BRPI0412890B8 (en) 2003-07-24 2021-05-25 Innate Pharma method of selecting an anti-kir2dl1 antibody or antigen-binding antibody fragment
US7754209B2 (en) 2003-07-26 2010-07-13 Trubion Pharmaceuticals Binding constructs and methods for use thereof
WO2005017529A1 (en) * 2003-07-29 2005-02-24 Genentech, Inc. Assay for human anti cd20 antibodies and uses therefor
CA2536408A1 (en) 2003-08-22 2005-03-03 Biogen Idec Ma Inc. Improved antibodies having altered effector function and methods for making the same
JP2007504138A (en) * 2003-08-29 2007-03-01 ジェネンテック・インコーポレーテッド Treatment of eye diseases
TWI333977B (en) * 2003-09-18 2010-12-01 Symphogen As Method for linking sequences of interest
CN102838675B (en) 2003-12-10 2014-07-30 梅达雷克斯有限责任公司 Ip-10 antibodies and their uses
US7741449B2 (en) 2003-12-10 2010-06-22 Medarex, Inc. Anti-interferon alpha antibodies
MXPA06006865A (en) * 2003-12-19 2006-08-23 Genentech Inc Detection of cd20 in transplant rejection.
AU2004305560A1 (en) * 2003-12-19 2005-07-07 Genentech, Inc. Detection of CD20 in therapy of autoimmune diseases
PT2311873T (en) 2004-01-07 2018-11-20 Novartis Vaccines & Diagnostics Inc M-csf-specific monoclonal antibody and uses thereof
DE602005018325D1 (en) 2004-02-19 2010-01-28 Genentech Inc ANTIBODIES WITH CORRECTED CDR
JP2007532680A (en) * 2004-04-16 2007-11-15 ジェネンテック・インコーポレーテッド Disease treatment method
RU2006140377A (en) * 2004-04-16 2008-05-27 Дженентек, Инк. (Us) METHOD FOR STRENGTHENING B-CELL DESTRUCTION
KR20070038453A (en) * 2004-04-16 2007-04-10 마크로제닉스, 인크. FCgamma;RIIB-SPECIFIC ANTIBODIES AND METHODS OF USE THEREOF
BRPI0510224A (en) * 2004-05-05 2007-10-23 Genentech Inc autoimmune disease prevention methods and industrialized article
WO2005110474A2 (en) * 2004-05-10 2005-11-24 Macrogenics, Inc. HUMANIZED FcϜRIIB SPECIFIC ANTIBODIES AND METHODS OF USE THEREOF
DE102004024617A1 (en) 2004-05-18 2005-12-29 Ganymed Pharmaceuticals Ag Differentially expressed in tumors gene products and their use
EP2583981A3 (en) 2004-05-28 2013-07-31 Agensys, Inc. Antibodies and related molecules that bind to PSCA proteins
BRPI0510915A (en) * 2004-06-04 2007-11-13 Genentech Inc Method for treating multiple sclerosis and manufactured article
PA8635501A1 (en) * 2004-06-04 2006-06-02 Genentech Inc USE OF AN ANTIBODY FOR THE TREATMENT OF LUPUS
EP2319526A1 (en) 2004-06-17 2011-05-11 Thrasos Therapeutics, Inc. Tdf-related compounds and analogs thereof
CA2570823C (en) 2004-06-21 2015-02-24 Medarex, Inc. Interferon alpha receptor 1 antibodies and their uses
BRPI0510674A (en) 2004-07-15 2007-12-26 Xencor Inc optimized fc variants
KR20070036187A (en) * 2004-07-22 2007-04-02 제넨테크, 인크. Method of treating sjogren's syndrome
KR20070047327A (en) 2004-07-26 2007-05-04 비오겐 아이덱 엠에이 아이엔씨. Anti-cd154 antibodies
AU2005270918B2 (en) 2004-08-03 2011-03-03 Innate Pharma S.A. Therapeutic and diagnostic methods and compositions targeting 4Ig-B7-H3 and its counterpart NK cell receptor
EP2213683B1 (en) 2004-08-04 2013-06-05 Mentrik Biotech, LLC Variant Fc regions
CA2577329A1 (en) * 2004-08-16 2006-03-02 Medimmune, Inc. Eph receptor fc variants with enhanced antibody dependent cell-mediated cytotoxicity activity
AU2005285347A1 (en) * 2004-08-19 2006-03-23 Genentech, Inc. Polypeptide variants with altered effector function
CA2580271A1 (en) * 2004-10-05 2006-04-20 Genentech, Inc. Method for treating vasculitis
JO3000B1 (en) 2004-10-20 2016-09-05 Genentech Inc Antibody Formulations.
EP1812068A4 (en) * 2004-10-29 2010-06-09 Medimmune Inc Methods of preventing and treating rsv infections and related conditions
US7632497B2 (en) * 2004-11-10 2009-12-15 Macrogenics, Inc. Engineering Fc Antibody regions to confer effector function
WO2007001457A2 (en) * 2004-11-12 2007-01-04 Xencor, Inc. Antibodies operably linked to selected chemoattractants
US20070135620A1 (en) * 2004-11-12 2007-06-14 Xencor, Inc. Fc variants with altered binding to FcRn
ES2396555T3 (en) * 2004-12-15 2013-02-22 Janssen Alzheimer Immunotherapy Antibodies that recognize beta amyloid peptide
JP2008523815A (en) * 2004-12-15 2008-07-10 エラン ファーマ インターナショナル リミテッド Humanized amyloid beta antibody for use in improving cognition
JP2008524247A (en) * 2004-12-15 2008-07-10 エラン ファーマ インターナショナル リミテッド Amyloid beta antibody for use in cognitive improvement
EP1674479A1 (en) * 2004-12-22 2006-06-28 Memorial Sloan-Kettering Cancer Center Modulation of Fc Gamma receptors for optimizing immunotherapy
EP1841454A4 (en) * 2005-01-13 2009-07-22 Genentech Inc Treatment method
CN102580084B (en) 2005-01-21 2016-11-23 健泰科生物技术公司 The fixed dosage of HER antibody is administered
DOP2006000029A (en) * 2005-02-07 2006-08-15 Genentech Inc ANTIBODY VARIANTS AND USES THEREOF. (VARIATIONS OF AN ANTIBODY AND USES OF THE SAME)
KR101289537B1 (en) 2005-02-15 2013-07-31 듀크 유니버시티 Anti-cd19 antibodies and uses in oncology
US20060263357A1 (en) 2005-05-05 2006-11-23 Tedder Thomas F Anti-CD19 antibody therapy for autoimmune disease
CN103251946A (en) 2005-02-23 2013-08-21 健泰科生物技术公司 Extending time to disease progression or survival in cancer patients using a her dimerization inhibitor
TW200714289A (en) * 2005-02-28 2007-04-16 Genentech Inc Treatment of bone disorders
US20060204505A1 (en) * 2005-03-08 2006-09-14 Sliwkowski Mark X Methods for identifying tumors responsive to treatment with HER dimerization inhibitors (HDIs)
EP2551282A3 (en) 2005-03-23 2013-02-13 Genmab A/S Antibodies against CD38 for treatment of multiple myeloma
AU2006230563B8 (en) 2005-03-31 2010-06-17 Agensys, Inc. Antibodies and related molecules that bind to 161P2F10B proteins
US9284375B2 (en) 2005-04-15 2016-03-15 Macrogenics, Inc. Covalent diabodies and uses thereof
US9963510B2 (en) * 2005-04-15 2018-05-08 Macrogenics, Inc. Covalent diabodies and uses thereof
US11254748B2 (en) 2005-04-15 2022-02-22 Macrogenics, Inc. Covalent diabodies and uses thereof
US9296816B2 (en) * 2005-04-15 2016-03-29 Macrogenics, Inc. Covalent diabodies and uses thereof
AR053579A1 (en) * 2005-04-15 2007-05-09 Genentech Inc TREATMENT OF INTESTINAL INFLAMMATORY DISEASE (IBD)
CN101203242A (en) * 2005-04-22 2008-06-18 健泰科生物技术公司 Method for treating dementia or alzheimer's disease by CD2O antibody
WO2006116260A2 (en) 2005-04-26 2006-11-02 Medimmune, Inc. Modulation of antibody effector function by hinge domain engineering
CN101228189A (en) * 2005-05-09 2008-07-23 格黎卡特生物技术股份公司 Antigen binding molecules having modified FC regions and altered binding to FC receptors
DK2161336T4 (en) 2005-05-09 2017-04-24 Ono Pharmaceutical Co Human monoclonal antibodies for programmed death 1 (PD-1) and methods for treating cancer using anti-PD-1 antibodies alone or in combination with other immunotherapies
KR20080046135A (en) * 2005-05-20 2008-05-26 제넨테크, 인크. Pretreatment of a biological sample from an autoimmune disease subject
US20070110757A1 (en) 2005-06-23 2007-05-17 Ziping Wei Antibody formulations having optimized aggregation and fragmentation profiles
AU2006265108C1 (en) 2005-07-01 2013-01-17 E. R. Squibb & Sons, L.L.C. Human monoclonal antibodies to programmed death ligand 1 (PD-L1)
US8309690B2 (en) * 2005-07-01 2012-11-13 Medimmune, Llc Integrated approach for generating multidomain protein therapeutics
WO2007008943A2 (en) 2005-07-08 2007-01-18 Xencor, Inc. Optimized anti-ep-cam antibodies
AU2006272597A1 (en) 2005-07-25 2007-02-01 Emergent Product Development Seattle Llc Single dose use of CD20-specific binding molecules
US20080279850A1 (en) * 2005-07-25 2008-11-13 Trubion Pharmaceuticals, Inc. B-Cell Reduction Using CD37-Specific and CD20-Specific Binding Molecules
EP1913028B1 (en) * 2005-07-28 2015-03-04 Novartis AG Use of antibody to m-csf
JP5457671B2 (en) * 2005-07-28 2014-04-02 ノバルティス アーゲー M-CSF specific monoclonal antibody and use thereof
DK1919503T3 (en) 2005-08-10 2014-12-15 Macrogenics Inc Identification and preparation of antibodies with variant fc regions and methods of use thereof
EP1762575A1 (en) 2005-09-12 2007-03-14 Ganymed Pharmaceuticals AG Identification of tumor-associated antigens for diagnosis and therapy
SI2497780T1 (en) 2005-09-20 2015-08-31 Thrasos Innovation, Inc. TDF-related compounds and analogs thereof
KR20080073293A (en) 2005-10-14 2008-08-08 메디뮨 엘엘씨 Cell display of antibody libraries
JP5006330B2 (en) 2005-10-21 2012-08-22 ノバルティス アーゲー Human antibodies against IL13 and therapeutic uses
US20070099246A1 (en) * 2005-11-03 2007-05-03 Sandy John D Antibodies, assays and kits to quantitate cartilage destruction
MY149159A (en) 2005-11-15 2013-07-31 Hoffmann La Roche Method for treating joint damage
CA2629306A1 (en) * 2005-11-23 2007-05-31 Genentech, Inc. Methods and compositions related to b cell assays
EP1790664A1 (en) 2005-11-24 2007-05-30 Ganymed Pharmaceuticals AG Monoclonal antibodies against claudin-18 for treatment of cancer
CN105859886A (en) 2005-12-02 2016-08-17 健泰科生物技术公司 Compositions and methods associated with antibodies that bind to IL-22 and IL-22R
CN103204931B (en) 2005-12-08 2016-08-03 米德列斯公司 The human monoclonal antibodies of anti-fucosyl-GM 1 and the method for use anti-fucosyl-GM 1 antibody
DK1973950T3 (en) 2006-01-05 2014-12-15 Genentech Inc ANTI-EphB4 ANTIBODIES AND METHODS FOR USING SAME
DK2463305T3 (en) 2006-01-12 2016-08-29 Alexion Pharma Inc Antibodies to OX-2 / CD200 and uses thereof
JP5368110B2 (en) 2006-01-20 2013-12-18 ジェネンテック， インコーポレイテッド Anti-ephrin B2 antibody and method of use thereof
AU2007212147A1 (en) * 2006-02-03 2007-08-16 Medimmune, Llc Protein formulations
EP2540741A1 (en) 2006-03-06 2013-01-02 Aeres Biomedical Limited Humanized anti-CD22 antibodies and their use in treatment of oncology, transplantation and autoimmune disease
CA2644903A1 (en) * 2006-03-10 2007-09-20 Macrogenics, Inc. Identification and engineering of antibodies with variant heavy chains and methods of using same
AR059851A1 (en) 2006-03-16 2008-04-30 Genentech Inc ANTIBODIES OF EGFL7 AND METHODS OF USE
US8278421B2 (en) 2006-03-20 2012-10-02 Xoma Techolology Ltd. Human antibodies specific for gastrin materials and methods
US20090220504A1 (en) 2006-03-21 2009-09-03 Anan Chuntharapai Combinatorial therapy
US8784810B2 (en) * 2006-04-18 2014-07-22 Janssen Alzheimer Immunotherapy Treatment of amyloidogenic diseases
ES2489646T3 (en) 2006-05-26 2014-09-02 Macrogenics, Inc. Humanized antibodies specific to Fc gamma RIIB and its methods of use
RS53168B (en) 2006-05-30 2014-06-30 Genentech Inc. Antibodies and immunoconjugates and uses therefor
CL2007001623A1 (en) * 2006-06-06 2008-01-18 Genentech Inc Anti-dll4 antibody; polynucleotide that encodes it; vector and host cell comprising said polynucleotide; method for making the antibody and immunojugate; method of detection of dll4 and diagnostic method of a disorder associated with dll4; composition comprising the antibody.
TW200817435A (en) * 2006-06-06 2008-04-16 Genentech Inc Compositions and methods for modulating vascular development
EP2032159B1 (en) * 2006-06-26 2015-01-07 MacroGenics, Inc. Combination of fcgammariib antibodies and cd20-specific antibodies and methods of use thereof
EP2505209A1 (en) 2006-06-26 2012-10-03 MacroGenics, Inc. Fcgamma-RIIB-specific antibodies and methods of the use thereof
AU2007275654A1 (en) 2006-07-19 2008-01-24 The Trustees Of The University Of Pennsylvania WSX-1/p28 as a target for anti-inflammatory responses
CN101626783A (en) * 2006-08-04 2010-01-13 诺华有限公司 EPHB3-specific antibody and its application
EP3415532A1 (en) 2006-08-18 2018-12-19 XOMA Technology Ltd. Prlr-specific antibody and uses thereof
EP2074144A4 (en) 2006-09-05 2011-03-16 Medarex Inc Antibodies to bone morphogenic proteins and receptors therefor and methods for their use
DK2066349T3 (en) 2006-09-08 2012-07-09 Medimmune Llc HUMANIZED ANTI-CD19 ANTIBODIES AND USE THEREOF IN TREATMENT OF TUMORS, TRANSPLANTATION AND AUTOIMMUNE DISEASES
EP2061906B1 (en) 2006-09-12 2011-08-31 Genentech, Inc. Methods and compositions for the diagnosis and treatment of lung cancer using pdgfra, kit or kdr gene as genetic marker
PL2486941T3 (en) 2006-10-02 2017-08-31 E. R. Squibb & Sons, L.L.C. Human antibodies that bind CXCR4 and uses thereof
US20080112961A1 (en) * 2006-10-09 2008-05-15 Macrogenics, Inc. Identification and Engineering of Antibodies with Variant Fc Regions and Methods of Using Same
US8613925B2 (en) 2006-10-19 2013-12-24 Csl Limited Anti-IL-13Rα1 antibodies and their uses thereof
WO2008060813A2 (en) 2006-10-19 2008-05-22 Merck & Co., Inc. High affinity antibody antagonists of interleukin-13 receptor alpha 1
RS53942B1 (en) 2006-10-27 2015-08-31 Genentech Inc. Antibodies and immunoconjugates and uses therefor
US8618248B2 (en) 2006-10-31 2013-12-31 President And Fellows Of Harvard College Phosphopeptide compositions and anti-phosphopeptide antibody compositions and methods of detecting phosphorylated peptides
CN111518218A (en) 2006-11-02 2020-08-11 阿塞勒隆制药公司 ALK1 receptor and ligand antagonists and uses thereof
MX2009005189A (en) 2006-11-15 2009-06-30 Medarex Inc Human monoclonal antibodies to btla and methods of use.
CA2671264C (en) 2006-11-30 2015-11-24 Research Development Foundation Improved immunoglobulin libraries
TWI447124B (en) 2006-12-01 2014-08-01 Medarex Llc Human antibodies that bind cd22 and uses thereof
WO2008070780A1 (en) 2006-12-07 2008-06-12 Novartis Ag Antagonist antibodies against ephb3
US8652466B2 (en) * 2006-12-08 2014-02-18 Macrogenics, Inc. Methods for the treatment of disease using immunoglobulins having Fc regions with altered affinities for FcγRactivating and FcγRinhibiting
CL2007003622A1 (en) 2006-12-13 2009-08-07 Medarex Inc Human anti-cd19 monoclonal antibody; composition comprising it; and tumor cell growth inhibition method.
KR20090088946A (en) 2006-12-14 2009-08-20 메다렉스, 인코포레이티드 Human antibodies that bind cd70 and uses thereof
TW200833711A (en) * 2006-12-22 2008-08-16 Genentech Inc Antibodies to insulin-like growth factor receptor
EP2125013A4 (en) * 2007-01-26 2010-04-07 Bioinvent Int Ab Dll4 signaling inhibitors and uses thereof
BRPI0806403A2 (en) 2007-02-09 2011-09-06 Genentech Inc anti-robo4 antibody, use an antibody and imaging method
RU2459868C2 (en) * 2007-03-01 2012-08-27 Симфоген А/С Method of cloning cognate antibodies
WO2008109440A2 (en) 2007-03-02 2008-09-12 Genentech, Inc. Predicting response to a her dimerisation inhibitor based on low her3 expression
WO2008112192A2 (en) 2007-03-08 2008-09-18 Kalobios Pharmaceuticals, Inc. Epha3 antibodies for the treatment of solid tumors
EP1970384A1 (en) * 2007-03-14 2008-09-17 Ganymed Pharmaceuticals AG Monoclonal antibodies for treatment of cancer
MX2009010120A (en) 2007-03-22 2009-10-19 Ucb Pharma Sa Binding proteins, including antibodies, antibody derivatives and antibody fragments, that specifically bind cd154 and uses thereof.
WO2008118324A2 (en) 2007-03-26 2008-10-02 Macrogenics, Inc. Composition and method of treating cancer with an anti-uroplakin ib antibody
US8003097B2 (en) * 2007-04-18 2011-08-23 Janssen Alzheimer Immunotherapy Treatment of cerebral amyloid angiopathy
AU2008242648B2 (en) * 2007-04-18 2013-09-12 Janssen Alzheimer Immunotherapy Prevention and treatment of cerebral amyloid angiopathy
WO2008134046A1 (en) 2007-04-27 2008-11-06 Genentech, Inc. Potent, stable and non-immunosuppressive anti-cd4 antibodies
EP2703011A3 (en) 2007-05-07 2014-03-26 MedImmune, LLC Anti-icos antibodies and their use in treatment of oncology, transplantation and autoimmune disease
EP1997832A1 (en) 2007-05-29 2008-12-03 Ganymed Pharmaceuticals AG Monoclonal antibodies against Claudin-18 for treatment of cancer
EP1997830A1 (en) 2007-06-01 2008-12-03 AIMM Therapeutics B.V. RSV specific binding molecules and means for producing them
PE20090321A1 (en) 2007-06-04 2009-04-20 Genentech Inc ANTI-NOTCH1 NRR ANTIBODIES, METHOD OF PREPARATION AND PHARMACEUTICAL COMPOSITION
ES2583377T3 (en) 2007-06-08 2016-09-20 Genentech, Inc. Gene expression markers of tumor resistance to HER2 inhibitor treatment
CA2691434C (en) 2007-06-21 2020-07-21 Macrogenics, Inc. Covalent diabodies and uses thereof
US20110077383A1 (en) * 2007-07-03 2011-03-31 Medimmune, Llc Hinge domain engineering
ES2751022T3 (en) 2007-07-09 2020-03-30 Genentech Inc Prevention of reduction of disulfide bonds during recombinant production of polypeptides
PL2474557T3 (en) 2007-07-16 2015-02-27 Genentech Inc Anti-CD79b antibodies and immunoconjugates and methods of use
US9845355B2 (en) 2007-07-16 2017-12-19 Genentech, Inc. Humanized anti-CD79b antibodies and immunoconjugates and methods of use
JP5889529B2 (en) 2007-07-27 2016-03-22 ヤンセン・サイエンシズ・アイルランド・ユーシー Treatment of amyloidogenic diseases
CA2694488A1 (en) 2007-07-31 2009-02-05 Medimmune, Llc Multispecific epitope binding proteins and uses thereof
DK2769729T3 (en) 2007-09-04 2019-04-23 Compugen Ltd POLYPEPTIDES AND POLYNUCLEOTIDES AND APPLICATIONS THEREOF AS A PHARMACEUTICAL OBJECTIVE FOR THE PRODUCTION OF MEDICINAL PRODUCTS AND BIOTECHNOLOGICAL PRODUCTS
MX342551B (en) 2007-09-26 2016-10-04 Chugai Pharmaceutical Co Ltd Modified antibody constant region.
TWI489993B (en) 2007-10-12 2015-07-01 Novartis Ag Compositions and methods of use for antibodies against sclerostin
DK2233149T3 (en) 2007-10-16 2016-05-17 Zymogenetics Inc COMBINATION OF TRANSMEMBRANAKTIVATOR AND CALCIUM MODULATOR AND cyclophilin-LIGAND INTERAKTOR (TACI) AND ANTI-CD20 MEANS FOR TREATMENT OF AUTO-IMMUNE DISEASE
JO3076B1 (en) * 2007-10-17 2017-03-15 Janssen Alzheimer Immunotherap Immunotherapy regimes dependent on apoe status
EP2567709B1 (en) 2007-11-02 2017-12-27 Novartis AG Molecules and methods for modulating low-density-lipoprotein receptor-related protein 6 (LRP6)
CN104888193A (en) 2007-11-07 2015-09-09 健泰科生物技术公司 Compositions and methods for treatment of microbial disorders
TWI468417B (en) 2007-11-30 2015-01-11 Genentech Inc Anti-vegf antibodies
CN110698561A (en) 2007-12-14 2020-01-17 诺沃—诺迪斯克有限公司 Anti-human NKG2D antibodies and uses thereof
WO2009117030A2 (en) * 2007-12-19 2009-09-24 Macrogenics, Inc. Improved compositions for the prevention and treatment of smallpox
US8092804B2 (en) 2007-12-21 2012-01-10 Medimmune Limited Binding members for interleukin-4 receptor alpha (IL-4Rα)-173
MX343594B (en) 2007-12-21 2016-11-11 Medimmune Ltd BINDING MEMBERS FOR INTERLEUKIN-4 RECEPTOR ALPHA (IL-4Ra) - 173.
US7914785B2 (en) 2008-01-02 2011-03-29 Bergen Teknologieverforing As B-cell depleting agents, like anti-CD20 antibodies or fragments thereof for the treatment of chronic fatigue syndrome
EP2077281A1 (en) 2008-01-02 2009-07-08 Bergen Teknologioverforing AS Anti-CD20 antibodies or fragments thereof for the treatment of chronic fatigue syndrome
TWI472339B (en) 2008-01-30 2015-02-11 Genentech Inc Composition comprising antibody that binds to domain ii of her2 and acidic variants thereof
RU2553566C2 (en) 2008-01-31 2015-06-20 Дженентек, Инк. ANTI-CD79b ANTIBODIES AND IMMUNOCONJUGATES AND METHODS FOR USING THEM
AU2009211340B2 (en) * 2008-02-04 2013-09-12 Lay Line Genomics S.P.A. Anti-TrkA antibodies and derivatives thereof
BRPI0906387A2 (en) 2008-02-05 2015-07-07 Bristol Myers Squibb Co Alpha 5 - beta 1 antibodies and their uses
AU2009212273B2 (en) * 2008-02-08 2014-07-31 Astrazeneca Ab Anti-IFNAR1 antibodies with reduced Fc ligand affinity
SG2014014138A (en) * 2008-03-31 2014-07-30 Genentech Inc Compositions and methods for treating and diagnosing asthma
DK2247304T3 (en) 2008-04-02 2016-09-26 Macrogenics Inc Her2 / neu-specific antibodies and methods of use thereof
MX2010010387A (en) 2008-04-02 2010-10-15 Macrogenics Inc Bcr-complex-specific antibodies and methods of using same.
CN105963695A (en) 2008-05-02 2016-09-28 阿塞勒隆制药公司 Methods and compositions based on ALK1 antagonists for modulating angiogenesis and pericyte coverage
US20110081347A1 (en) * 2008-06-04 2011-04-07 Macrogenics, Inc. Antibodies with Altered Binding to FcRn and Methods of Using Same
CA2731956A1 (en) 2008-07-25 2010-01-28 Daniel S. Gareau Rapid confocal microscopy to support surgical procedures
MY159396A (en) 2008-08-05 2016-12-30 Novartis Ag Compositions and methods for antibodies targeting complement protein c5
AR072999A1 (en) 2008-08-11 2010-10-06 Medarex Inc HUMAN ANTIBODIES THAT JOIN GEN 3 OF LYMPHOCYTARY ACTIVATION (LAG-3) AND THE USES OF THESE
BRPI0917352A2 (en) * 2008-08-29 2017-08-22 Symphogen As METHODS FOR PRODUCING A LIBRARY OF COGNATE PAIRS COMPRISING LINKED SEQUENCES ENCODING VARIABLE REGIONS AND FOR GENERATING A VECTOR ENCODING A CHIMERIC ANTIBODY WITH CONSTANT HUMAN REGIONS AND NON-HUMAN VARIABLE REGIONS, VECTOR LIBRARY, AND METHOD FOR PRODUCING A LIBRARY OF AVIAN-DERIVED IMMUNOGLOBULIN SEQUENCES
EP2166021A1 (en) 2008-09-16 2010-03-24 Ganymed Pharmaceuticals AG Monoclonal antibodies for treatment of cancer
AR073295A1 (en) 2008-09-16 2010-10-28 Genentech Inc METHODS TO TREAT PROGRESSIVE MULTIPLE SCLEROSIS. MANUFACTURING ARTICLE.
WO2010032060A1 (en) 2008-09-19 2010-03-25 Medimmune Llc Antibodies directed to dll4 and uses thereof
CN110317272A (en) 2008-10-14 2019-10-11 霍夫曼-拉罗奇有限公司 Immunoglobulin variants and application thereof
US9067981B1 (en) 2008-10-30 2015-06-30 Janssen Sciences Ireland Uc Hybrid amyloid-beta antibodies
US8298533B2 (en) 2008-11-07 2012-10-30 Medimmune Limited Antibodies to IL-1R1
US20110293605A1 (en) 2008-11-12 2011-12-01 Hasige Sathish Antibody formulation
CN102271708A (en) * 2008-11-13 2011-12-07 新兴产品开发西雅图有限公司 CD37 immunotherapeutic combination therapies and uses thereof
HUE032693T2 (en) 2008-11-22 2017-10-30 Hoffmann La Roche Use of anti-vegf antibody in combination with chemotherapy for treating breast cancer
WO2010063785A2 (en) 2008-12-03 2010-06-10 Genmab A/S Antibody variants having modifications in the constant region
EP2865689A1 (en) 2008-12-08 2015-04-29 Compugen Ltd. FAM26F polypeptides and polynucleotides, and uses thereof as a drug target for producing drugs and biologics
SG10201708690SA (en) 2008-12-09 2017-12-28 Genentech Inc Anti-pd-l1 antibodies and their use to enhance t-cell function
US8775090B2 (en) 2008-12-12 2014-07-08 Medimmune, Llc Crystals and structure of a human IgG Fc variant with enhanced FcRn binding
CN106220734A (en) 2008-12-19 2016-12-14 宏观基因有限公司 Covalent diabodies and application thereof
WO2010075249A2 (en) 2008-12-22 2010-07-01 Genentech, Inc. A method for treating rheumatoid arthritis with b-cell antagonists
KR20170143025A (en) 2008-12-23 2017-12-28 제넨테크, 인크. Immunoglobulin variants with altered binding to protein a
JP2012513194A (en) 2008-12-23 2012-06-14 アストラゼネカ アクチボラグ Targeted binding agents directed to α5β1 and uses thereof
WO2010078526A1 (en) 2008-12-31 2010-07-08 Biogen Idec Ma Inc. Anti-lymphotoxin antibodies
US20110142836A1 (en) * 2009-01-02 2011-06-16 Olav Mella B-cell depleting agents for the treatment of chronic fatigue syndrome
HRP20231525T1 (en) 2009-02-20 2024-03-15 Astellas Pharma Inc. Methods and compositions for diagnosis and treatment of cancer
WO2010095031A2 (en) * 2009-02-23 2010-08-26 Glenmark Pharmaceuticals S.A. Humanized antibodies that bind to cd19 and their uses
AR075715A1 (en) * 2009-03-05 2011-04-20 Novartis Ag FORMULATION OF LIOFILIZED ANTIBODY
MX2011009220A (en) 2009-03-05 2011-09-28 Medarex Inc Fully human antibodies specific to cadm1.
CA2753995A1 (en) 2009-03-06 2010-09-10 Kalobios Pharmaceuticals, Inc. Treatment of leukemias and chronic myeloproliferative diseases with antibodies to epha3
MA33198B1 (en) 2009-03-20 2012-04-02 Genentech Inc ANTI-HER DI-SPECIFIC ANTIBODIES
US8124740B2 (en) 2009-03-25 2012-02-28 Genentech, Inc. Anti- α5 β1 antibodies and uses thereof
CN102378767B (en) 2009-03-25 2015-01-14 健泰科生物技术公司 Anti-fgfr3 antibodies and methods using same
NZ595574A (en) 2009-04-01 2013-11-29 Genentech Inc Anti-fcrh5 antibodies and immunoconjugates and methods of use
US20100256340A1 (en) 2009-04-07 2010-10-07 Ulrich Brinkmann Trivalent, bispecific antibodies
US20100297127A1 (en) 2009-04-08 2010-11-25 Ghilardi Nico P Use of il-27 antagonists to treat lupus
EA030182B1 (en) 2009-04-20 2018-07-31 Оксфорд Байотерепьютикс Лтд. Antibodies specific to cadherin-17
WO2010124113A1 (en) 2009-04-23 2010-10-28 Infinity Pharmaceuticals, Inc. Anti-fatty acid amide hydrolase-2 antibodies and uses thereof
EP3275900A1 (en) 2009-04-27 2018-01-31 Novartis AG Compositions and methods for increasing muscle growth
EP2424894A1 (en) 2009-04-27 2012-03-07 Novartis AG Composition and methods of use for therapeutic antibodies specific for the il-12 receptore betal subunit
DK2429574T3 (en) 2009-05-15 2015-06-22 Univ Health Network COMPOSITIONS AND METHODS FOR TREATING HEMATOLOGICAL CANCERS TARGETING THE SIRP-CD47 INTERACTION
US9676845B2 (en) 2009-06-16 2017-06-13 Hoffmann-La Roche, Inc. Bispecific antigen binding proteins
PE20121494A1 (en) 2009-06-18 2012-11-01 Pfizer ANTI NOTCH-1 ANTIBODIES
AU2010273585B2 (en) 2009-07-13 2015-04-23 Genentech, Inc. Diagnostic methods and compositions for treatment of cancer
TW201106972A (en) 2009-07-27 2011-03-01 Genentech Inc Combination treatments
SG177689A1 (en) 2009-07-31 2012-02-28 Organon Nv Fully human antibodies to btla
SG178177A1 (en) 2009-07-31 2012-03-29 Genentech Inc Inhibition of tumor metastasis using bv8- or g-csf-antagonists
LT2464725T (en) 2009-08-11 2020-06-10 F. Hoffmann-La Roche Ag Production of proteins in glutamine-free cell culture media
MX2012001716A (en) 2009-08-14 2012-04-02 Genentech Inc Biological markers for monitoring patient response to vegf antagonists.
TW201431558A (en) 2009-08-15 2014-08-16 建南德克公司 Anti-angiogenesis therapy for the treatment of previously treated breast cancer
WO2011021146A1 (en) 2009-08-20 2011-02-24 Pfizer Inc. Osteopontin antibodies
CA2772051C (en) 2009-08-24 2020-08-18 Amunix Operating Inc. Coagulation factor ix compositions and methods of making and using same
DK2473522T3 (en) 2009-09-02 2016-11-28 Genentech Inc Smoothened MUTANT AND METHODS OF USING THE SAME
MX353186B (en) 2009-09-03 2018-01-05 Genentech Inc Methods for treating, diagnosing, and monitoring rheumatoid arthritis.
WO2011029823A1 (en) 2009-09-09 2011-03-17 Novartis Ag Monoclonal antibody reactive with cd63 when expressed at the surface of degranulated mast cells
CA2773662A1 (en) 2009-09-11 2011-03-17 Genentech, Inc. Method to identify a patient with an increased likelihood of responding to an anti-cancer agent
NZ701769A (en) 2009-09-16 2016-06-24 Genentech Inc Coiled coil and/or tether containing protein complexes and uses thereof
ES2530732T3 (en) 2009-09-17 2015-03-05 Hoffmann La Roche Diagnostic procedures for lung cancer
EP2480888B1 (en) 2009-09-25 2016-11-30 XOMA Technology Ltd. Screening methods
US8926976B2 (en) 2009-09-25 2015-01-06 Xoma Technology Ltd. Modulators
US8568726B2 (en) 2009-10-06 2013-10-29 Medimmune Limited RSV specific binding molecule
EP2486141B1 (en) 2009-10-07 2018-01-10 MacroGenics, Inc. Fc region-containing polypeptides that exhibit improved effector function due to alterations of the extent of fucosylation, and methods for their use
US20120231004A1 (en) 2009-10-13 2012-09-13 Oxford Biotherapeutic Ltd. Antibodies
PL2488554T3 (en) 2009-10-14 2020-03-31 Humanigen, Inc. Antibodies to epha3
KR20120105447A (en) 2009-10-22 2012-09-25 제넨테크, 인크. Anti-hepsin antibodies and methods using same
WO2011056502A1 (en) 2009-10-26 2011-05-12 Genentech, Inc. Bone morphogenetic protein receptor type ii compositions and methods of use
WO2011056494A1 (en) 2009-10-26 2011-05-12 Genentech, Inc. Activin receptor-like kinase-1 antagonist and vegfr3 antagonist combinations
WO2011056497A1 (en) 2009-10-26 2011-05-12 Genentech, Inc. Activin receptor type iib compositions and methods of use
WO2011053982A2 (en) 2009-11-02 2011-05-05 University Of Washington Therapeutic nuclease compositions and methods
CN102782149B (en) 2009-11-04 2014-11-12 默沙东公司 Engineered anti-TSLP antibody
ES2636971T3 (en) 2009-11-05 2017-10-10 F. Hoffmann-La Roche Ag Procedures and composition for secretion of heterogeneous polypeptides
EP3689911A1 (en) 2009-11-11 2020-08-05 Ganymed Pharmaceuticals GmbH Antibodies specific for claudin 6 (cldn6)
EP2322555A1 (en) 2009-11-11 2011-05-18 Ganymed Pharmaceuticals AG Antibodies specific for claudin 6 (CLDN6)
NZ628923A (en) 2009-11-24 2016-02-26 Medimmune Ltd Targeted binding agents against b7-h1
EP2327725A1 (en) 2009-11-26 2011-06-01 InflaRx GmbH Anti-C5a binding moieties with high blocking activity
US9428586B2 (en) 2009-12-01 2016-08-30 Compugen Ltd Heparanase splice variant
US8722615B2 (en) 2009-12-02 2014-05-13 Acceleron Pharma, Inc. Compositions and methods for increasing serum half-life
LT2510012T (en) 2009-12-09 2017-07-25 Bayer Pharma Aktiengesellschaft Anti-c4.4a antibodies and uses thereof
TWI505836B (en) 2009-12-11 2015-11-01 Genentech Inc Anti-vegf-c antibodies and methods using same
MX2012007318A (en) 2009-12-22 2012-07-20 Novartis Ag Tetravalent cd47-antibody constant region fusion protein for use in therapy.
PL2516465T3 (en) 2009-12-23 2016-11-30 Anti-bv8 antibodies and uses thereof
CN102906115A (en) 2010-01-11 2013-01-30 阿雷克森制药公司 Biomarkers of immunomodulatory effects in humans treated with anti-cd200 antibodies
US8609092B2 (en) 2010-02-08 2013-12-17 Agensys, Inc. Antibody drug conjugates (ADC) that bind to 161P2F10B proteins
RU2573994C2 (en) 2010-02-10 2016-01-27 Иммьюноджен, Инк Anti-cd20 antibodies and thereof application
EA201201113A1 (en) 2010-02-10 2013-03-29 Новартис Аг METHODS AND CONNECTIONS FOR GROWTH OF MUSCLE
MX2012008958A (en) 2010-02-18 2012-08-23 Genentech Inc Neuregulin antagonists and use thereof in treating cancer.
KR20180028561A (en) 2010-02-23 2018-03-16 제넨테크, 인크. Anti-angiogenesis therapy for the treatment of ovarian cancer
US8802091B2 (en) 2010-03-04 2014-08-12 Macrogenics, Inc. Antibodies reactive with B7-H3 and uses thereof
NZ705128A (en) 2010-03-04 2015-04-24 Macrogenics Inc Antibodies reactive with b7-h3, immunologically active fragments thereof and uses thereof
EP2371864A1 (en) 2010-03-23 2011-10-05 Ganymed Pharmaceuticals AG Monoclonal antibodies for treatment of cancer
BR112012022044A2 (en) 2010-03-24 2020-08-25 Genentech Inc ''antibody, immunoconjugate, pharmaceutical formulation, antibody use, treatment method, isolated bispecific antibody and host cell''.
US10745467B2 (en) 2010-03-26 2020-08-18 The Trustees Of Dartmouth College VISTA-Ig for treatment of autoimmune, allergic and inflammatory disorders
TW201138821A (en) 2010-03-26 2011-11-16 Roche Glycart Ag Bispecific antibodies
US20150231215A1 (en) 2012-06-22 2015-08-20 Randolph J. Noelle VISTA Antagonist and Methods of Use
CN103119054B (en) 2010-03-26 2017-01-18 达特茅斯大学理事会 VISTA regulatory T cell mediator protein, VISTA binding agents and use thereof
CA2794745A1 (en) 2010-03-29 2011-10-06 Zymeworks, Inc. Antibodies with enhanced or suppressed effector function
WO2011133931A1 (en) 2010-04-22 2011-10-27 Genentech, Inc. Use of il-27 antagonists for treating inflammatory bowel disease
PL2563813T3 (en) 2010-04-30 2016-01-29 Alexion Pharma Inc Anti-c5a antibodies and methods for using the antibodies
CA2798390A1 (en) 2010-05-06 2011-11-10 Novartis Ag Compositions and methods of use for therapeutic low density lipoprotein - related protein 6 (lrp6) multivalent antibodies
SG185416A1 (en) 2010-05-06 2012-12-28 Novartis Ag Compositions and methods of use for therapeutic low density lipoprotein -related protein 6 (lrp6) antibodies
WO2011146568A1 (en) 2010-05-19 2011-11-24 Genentech, Inc. Predicting response to a her inhibitor
WO2011147834A1 (en) 2010-05-26 2011-12-01 Roche Glycart Ag Antibodies against cd19 and uses thereof
WO2011153243A2 (en) 2010-06-02 2011-12-08 Genentech, Inc. Anti-angiogenesis therapy for treating gastric cancer
WO2011153224A2 (en) 2010-06-02 2011-12-08 Genentech, Inc. Diagnostic methods and compositions for treatment of cancer
AU2011261362B2 (en) 2010-06-03 2016-06-09 Genentech, Inc. Immuno-pet imaging of antibodies and immunoconjugates and uses therefor
CN103080136B (en) 2010-06-18 2015-08-12 霍夫曼-拉罗奇有限公司 Anti-Axl antibody and using method
WO2011161119A1 (en) 2010-06-22 2011-12-29 F. Hoffmann-La Roche Ag Antibodies against insulin-like growth factor i receptor and uses thereof
WO2011161189A1 (en) 2010-06-24 2011-12-29 F. Hoffmann-La Roche Ag Anti-hepsin antibodies and methods of use
EP2404936A1 (en) 2010-07-06 2012-01-11 Ganymed Pharmaceuticals AG Cancer therapy using CLDN6 target-directed antibodies in vivo
AU2011274423B2 (en) 2010-07-09 2016-02-11 Bioverativ Therapeutics Inc. Chimeric clotting factors
NZ605449A (en) 2010-07-09 2015-03-27 Genentech Inc Anti-neuropilin antibodies and methods of use
EP2409993A1 (en) 2010-07-19 2012-01-25 International-Drug-Development-Biotech Anti-CD19 antibody having ADCC function with improved glycosylation profile
EP2409712A1 (en) 2010-07-19 2012-01-25 International-Drug-Development-Biotech Anti-CD19 antibody having ADCC and CDC functions and improved glycosylation profile
WO2012010582A1 (en) 2010-07-21 2012-01-26 Roche Glycart Ag Anti-cxcr5 antibodies and methods of use
JP2013538191A (en) 2010-07-23 2013-10-10 トラスティーズ オブ ボストン ユニバーシティ Anti-DEsupR inhibitors as therapeutics for inhibition of pathological angiogenesis and tumor cell invasiveness and for molecular imaging and targeted delivery
EP2601216B1 (en) 2010-08-02 2018-01-03 MacroGenics, Inc. Covalent diabodies and uses thereof
BR112013002535A2 (en) 2010-08-03 2019-09-24 Hoffmann La Roche biomarkers of chronic lymphocytic leukemia (cll)
RU2013106217A (en) 2010-08-05 2014-09-10 Ф. Хоффманн-Ля Рош Аг HYBRID PROTEIN FROM ANTIBODIES AGAINST MHC AND ANTIVIRAL CYTOKINE
AU2011288412A1 (en) 2010-08-13 2013-02-21 Medimmune Limited Monomeric polypeptides comprising variant Fc regions and methods of use
AU2011288487B2 (en) 2010-08-13 2015-10-01 Roche Glycart Ag Anti-FAP antibodies and methods of use
MX2013001336A (en) 2010-08-13 2013-03-08 Roche Glycart Ag Anti-tenascin-c a2 antibodies and methods of use.
WO2012022734A2 (en) 2010-08-16 2012-02-23 Medimmune Limited Anti-icam-1 antibodies and methods of use
CN103080134B (en) 2010-08-20 2015-11-25 诺华股份有限公司 The antibody of EGF-R ELISA 3 (HER3)
CN103068846B9 (en) 2010-08-24 2016-09-28 弗·哈夫曼-拉罗切有限公司 Bispecific antibodies comprising disulfide-stabilized Fv fragments
EP2609117B1 (en) 2010-08-25 2015-09-16 F.Hoffmann-La Roche Ag Antibodies against il-18r1 and uses thereof
DK2612151T3 (en) 2010-08-31 2017-10-02 Genentech Inc BIOMARKETS AND METHODS OF TREATMENT
WO2012028683A1 (en) 2010-09-02 2012-03-08 Novartis Ag Antibody gel system for sustained drug delivery
CN103328511B (en) 2010-09-10 2016-01-20 埃派斯进有限公司 Anti-IL-1 β antibody and using method thereof
US8999335B2 (en) 2010-09-17 2015-04-07 Compugen Ltd. Compositions and methods for treatment of drug resistant multiple myeloma
KR20130130709A (en) 2010-09-29 2013-12-02 어젠시스 인코포레이티드 Antibody drug conjugates (adc) that bind to 191p4d12 proteins
JP2013543384A (en) 2010-10-05 2013-12-05 ノバルティス アーゲー Anti-IL12Rbeta1 antibody and its use in the treatment of autoimmune and inflammatory diseases
AU2011312205B2 (en) 2010-10-05 2015-08-13 Curis, Inc. Mutant smoothened and methods of using the same
AU2011325833C1 (en) 2010-11-05 2017-07-13 Zymeworks Bc Inc. Stable heterodimeric antibody design with mutations in the Fc domain
JP6163429B2 (en) 2010-11-10 2017-07-12 ジェネンテック， インコーポレイテッド Methods and compositions for neurological disease immunotherapy
US10208129B2 (en) 2010-12-06 2019-02-19 National Research Council Of Canada Antibodies selective for cells presenting ErbB2 at high density
AU2011342799B2 (en) 2010-12-15 2016-06-09 Wyeth Llc Anti-Notch1 antibodies
NZ609493A (en) 2010-12-16 2015-11-27 Genentech Inc Diagnosis and treatments relating to th2 inhibition
EA028744B1 (en) 2010-12-20 2017-12-29 Дженентек, Инк. Anti-mesothelin antibodies and immunoconjugates
KR20130118925A (en) 2010-12-22 2013-10-30 제넨테크, 인크. Anti-pcsk9 antibodies and methods of use
EP2655413B1 (en) 2010-12-23 2019-01-16 F.Hoffmann-La Roche Ag Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery
WO2012092539A2 (en) 2010-12-31 2012-07-05 Takeda Pharmaceutical Company Limited Antibodies to dll4 and uses thereof
ES2860748T3 (en) 2011-01-24 2021-10-05 Gilead Sciences Inc Selective Antibodies for EGFR-Presenting Cells with High Density
WO2012106634A1 (en) 2011-02-03 2012-08-09 Alexion Pharmaceuticals, Inc. Use of an anti-cd200 antibody for prolonging the survival of allografts
RU2013140975A (en) 2011-02-28 2015-04-10 Дженентек, Инк. BIOLOGICAL MARKERS AND METHODS FOR PREDICTING SUSCEPTIBILITY TO B-CELL ANTAGONISTS
CA2824824A1 (en) 2011-02-28 2012-09-07 F. Hoffmann-La Roche Ag Monovalent antigen binding proteins
CA2825081A1 (en) 2011-02-28 2012-09-07 Birgit Bossenmaier Antigen binding proteins
JP6097702B2 (en) 2011-03-03 2017-03-15 アペクシジェン， インコーポレイテッド Anti-IL-6 receptor antibody and method of use thereof
JP5832559B2 (en) 2011-03-10 2015-12-16 オメロス コーポレーション Generation of anti-FN14 monoclonal antibodies by accelerated antibody evolution ex vivo
RU2013143358A (en) 2011-04-07 2015-05-20 Дженентек, Инк. ANTI-FGFR4 ANTIBODIES AND WAYS OF THEIR APPLICATION
ES2608835T3 (en) 2011-04-13 2017-04-17 Bristol-Myers Squibb Company Fc fusion proteins comprising new linkers or arrangements
US9409987B2 (en) 2011-04-15 2016-08-09 Compugen Ltd Polypeptides and polynucleotides, and uses thereof for treatment of immune related disorders and cancer
CN108341863A (en) 2011-04-20 2018-07-31 阿塞勒隆制药公司 Endoglin polypeptide and application thereof
HUE037651T2 (en) 2011-04-20 2018-09-28 Medimmune Llc Antibodies and other molecules that bind b7-h1 and pd-1
BR112013027829B1 (en) 2011-04-29 2022-05-10 Apexigen, Inc Anti-cd40 antibodies or antigen-binding fragments thereof, a composition comprising said antibodies and use of said composition to treat or ameliorate symptoms of cancer, autoimmune disease or inflammatory disease
SI2704737T1 (en) 2011-04-29 2018-06-29 University Of Washington Therapeutic nuclease compositions and methods
WO2012146630A1 (en) 2011-04-29 2012-11-01 F. Hoffmann-La Roche Ag N-terminal acylated polypeptides, methods for their production and uses thereof
MX2013013054A (en) 2011-05-12 2014-02-20 Genentech Inc Multiple reaction monitoring lc-ms/ms method to detect therapeutic antibodies in animal samples using framework signature peptides.
RS58087B1 (en) 2011-05-13 2019-02-28 Ganymed Pharmaceuticals Gmbh Antibodies for treatment of cancer expressing claudin 6
DK2707391T3 (en) 2011-05-13 2018-02-05 Gamamabs Pharma ANTIBODIES AGAINST HER3
PE20140995A1 (en) 2011-05-16 2014-08-23 Genentech Inc FGFR1 AGONISTS AND THEIR METHODS OF USE
WO2012162160A1 (en) 2011-05-20 2012-11-29 Momenta Pharmaceuticals, Inc. Modified glycoproteins
EP2714733B1 (en) 2011-05-21 2019-01-23 MacroGenics, Inc. Cd3-binding molecules capable of binding to human and non-human cd3
AU2012259162C1 (en) 2011-05-21 2020-05-21 Macrogenics, Inc. Deimmunized serum-binding domains and their use for extending serum half-life
CA2837169C (en) 2011-05-24 2021-11-09 Zyngenia, Inc. Multispecific complexes comprising angiopoietin-2-binding peptide and their uses
SG10201902706VA (en) 2011-06-03 2019-04-29 Xoma Technology Ltd Antibodies specific for tgf-beta
US9561274B2 (en) 2011-06-07 2017-02-07 University Of Hawaii Treatment and prevention of cancer with HMGB1 antagonists
WO2012170740A2 (en) 2011-06-07 2012-12-13 University Of Hawaii Biomarker of asbestos exposure and mesothelioma
CA2838833A1 (en) 2011-06-10 2012-12-13 Biogen Idec Ma Inc. Pro-coagulant compounds and methods of use thereof
WO2012172495A1 (en) 2011-06-14 2012-12-20 Novartis Ag Compositions and methods for antibodies targeting tem8
BR112013032235A2 (en) 2011-06-15 2016-11-22 Hoffmann La Roche anti-human epo receptor antibodies and methods of use
WO2012172521A1 (en) 2011-06-16 2012-12-20 Novartis Ag Soluble proteins for use as therapeutics
MX354663B (en) 2011-06-22 2018-03-14 Hoffmann La Roche Removal of target cells by circulating virus-specific cytotoxic t-cells using mhc class i comprising complexes.
KR20140104944A (en) 2011-06-22 2014-08-29 인쎄름 (엥스띠뛰 나씨오날 드 라 쌍떼 에 드 라 흐쉐르슈 메디깔) Anti-axl antibodies and uses thereof
US9409988B2 (en) 2011-06-22 2016-08-09 Inserm (Institut National De La Sante Et De La Recherche Medicale) Anti-Axl antibodies and uses thereof
JP6247209B2 (en) 2011-06-28 2017-12-13 オックスフォード ビオトヘラペウトイクス エルティーディー． Antibody to ADP ribosyl cyclase 2
AR086823A1 (en) 2011-06-30 2014-01-22 Genentech Inc ANTI-C-MET ANTIBODY FORMULATIONS, METHODS
AU2012277376B2 (en) 2011-06-30 2016-11-24 Compugen Ltd. Polypeptides and uses thereof for treatment of autoimmune disorders and infection
PL2726099T3 (en) 2011-07-01 2018-12-31 Novartis Ag Method for treating metabolic disorders
CA3158257A1 (en) 2011-07-11 2013-01-17 Ichnos Sciences SA Antibodies that bind to ox40 and their uses
US9738707B2 (en) 2011-07-15 2017-08-22 Biogen Ma Inc. Heterodimeric Fc regions, binding molecules comprising same, and methods relating thereto
EP2731970B1 (en) 2011-07-15 2018-11-28 MorphoSys AG Antibodies that are cross-reactive for macrophage migration inhibitory factor (mif) and d-dopachrome tautomerase (d-dt)
US20130022551A1 (en) 2011-07-22 2013-01-24 Trustees Of Boston University DEspR ANTAGONISTS AND AGONISTS AS THERAPEUTICS
JP5944994B2 (en) 2011-08-12 2016-07-05 オメロス コーポレーション Anti-FZD10 monoclonal antibodies and methods for their use
JP6120848B2 (en) 2011-08-15 2017-04-26 メディミューン，エルエルシー Anti-B7-H4 antibody and use thereof
CA2842375A1 (en) 2011-08-17 2013-02-21 Erica Jackson Neuregulin antibodies and uses thereof
RU2014109985A (en) 2011-08-17 2015-09-27 Дженентек, Инк. INHIBITION OF ANGIOGENESIS IN REFRACTOR TUMORS
BR112014003999A2 (en) 2011-08-23 2017-06-13 Roche Glycart Ag isolated antibody that binds to a near epitope near the human mcsp membrane, isolated nucleic acid, host cell, method of producing an antibody, immunoconjugate, pharmaceutical formulation, use of the antibody, method of treating cancer patients, unduction method of cell lysis in individuals and mcsp immunohistochemical test
GB201212550D0 (en) 2012-07-13 2012-08-29 Novartis Ag B cell assay
UY34317A (en) 2011-09-12 2013-02-28 Genzyme Corp T cell antireceptor antibody (alpha) / ß
WO2013039954A1 (en) 2011-09-14 2013-03-21 Sanofi Anti-gitr antibodies
MX2014003094A (en) 2011-09-15 2014-04-25 Genentech Inc Methods of promoting differentiation.
CN103930111A (en) 2011-09-19 2014-07-16 霍夫曼-拉罗奇有限公司 Combination treatments comprising C-MET antagonists and B-RAF antagonists
BR112014007487A2 (en) 2011-09-30 2017-04-04 Dana Farber Cancer Inst Inc therapeutic peptides
US9663573B2 (en) 2011-10-05 2017-05-30 Genentech, Inc. Methods of treating liver conditions using Notch2 antagonists
CN104080471B (en) 2011-10-14 2018-08-10 诺华股份有限公司 Antibody and method for Wnt approach relevant diseases
US9738727B2 (en) 2011-10-14 2017-08-22 Genentech, Inc. Anti-HtrA1 antibodies and methods of use
WO2013056148A2 (en) 2011-10-15 2013-04-18 Genentech, Inc. Methods of using scd1 antagonists
WO2013059531A1 (en) 2011-10-20 2013-04-25 Genentech, Inc. Anti-gcgr antibodies and uses thereof
SG11201401815XA (en) 2011-10-28 2014-05-29 Genentech Inc Therapeutic combinations and methods of treating melanoma
AU2012332587B2 (en) 2011-11-01 2017-02-23 Bionomics, Inc. Antibodies and methods of treating cancer
AU2012332588B2 (en) 2011-11-01 2017-09-07 Bionomics, Inc. Methods of blocking cancer stem cell growth
EP2773667A1 (en) 2011-11-01 2014-09-10 Bionomics, Inc. Anti-gpr49 antibodies
EP2773664A1 (en) 2011-11-01 2014-09-10 Bionomics, Inc. Anti-gpr49 antibodies
WO2013067098A1 (en) 2011-11-02 2013-05-10 Apexigen, Inc. Anti-kdr antibodies and methods of use
KR102052774B1 (en) 2011-11-04 2019-12-04 자임워크스 인코포레이티드 Stable heterodimeric antibody design with mutations in the fc domain
US9580509B2 (en) 2011-11-07 2017-02-28 Medimmune, Llc Multispecific and multivalent binding proteins and uses thereof
RU2014124842A (en) 2011-11-21 2015-12-27 Дженентек, Инк. CLEANING ANTI-C-MET ANTIBODIES
RU2014124530A (en) 2011-11-23 2015-12-27 Идженика, Инк. ANTIBODIES TO CD98 AND WAYS OF THEIR APPLICATION
AR088941A1 (en) 2011-11-23 2014-07-16 Bayer Ip Gmbh ANTI-FGFR2 ANTIBODIES AND THEIR USES
US20130251703A1 (en) 2011-12-05 2013-09-26 Novartis Ag Antibodies for epidermal growth factor receptor 3 (her3) directed to domain iii and domain iv of her3
SG11201402739YA (en) 2011-12-05 2014-06-27 Novartis Ag Antibodies for epidermal growth factor receptor 3 (her3) directed to domain ii of her3
US9416179B2 (en) 2011-12-05 2016-08-16 X-Body, Inc. PDGF receptor beta binding polypeptides
EP2788024A1 (en) 2011-12-06 2014-10-15 F.Hoffmann-La Roche Ag Antibody formulation
RU2685867C2 (en) 2011-12-15 2019-04-23 Алтернатив Инновейтив Текнолоджиз Ллц Hybrid proteins and protein conjugates based on heat shock protein-70 (hsp70) and methods for use thereof (versions)
US9527927B2 (en) 2011-12-20 2016-12-27 Medimmune, Llc Modified polypeptides for bispecific antibody scaffolds
EP2794656B1 (en) 2011-12-21 2019-02-27 Novartis AG Compositions comprising antibodies targeting factor p and c5
AR089434A1 (en) 2011-12-23 2014-08-20 Genentech Inc PROCEDURE TO PREPARE FORMULATIONS WITH HIGH CONCENTRATION OF PROTEINS
WO2013101771A2 (en) 2011-12-30 2013-07-04 Genentech, Inc. Compositions and method for treating autoimmune diseases
CN104411717A (en) 2012-01-09 2015-03-11 斯克利普斯研究所 Humanized antibodies with ultralong CDR3S
AU2013208003B2 (en) 2012-01-09 2017-12-14 The Scripps Research Institute Ultralong complementarity determining regions and uses thereof
HUE046396T2 (en) 2012-01-12 2020-02-28 Bioverativ Therapeutics Inc Chimeric factor viii polypeptides and uses thereof
AU2013207778B2 (en) 2012-01-13 2017-10-12 Genentech, Inc. Biological markers for identifying patients for treatment with VEGF antagonists
JP6242813B2 (en) 2012-01-18 2017-12-06 ジェネンテック， インコーポレイテッド Anti-LRP5 antibody and method of use
WO2013109856A2 (en) 2012-01-18 2013-07-25 Genentech, Inc. Methods of using fgf19 modulators
TW201334789A (en) 2012-01-31 2013-09-01 Genentech Inc Anti-IgE antibodies and methods using same
IN2014KN00848A (en) 2012-02-01 2015-10-02 Compugen Ltd
MX2014009565A (en) 2012-02-10 2014-11-10 Genentech Inc Single-chain antibodies and other heteromultimers.
CN113398268A (en) 2012-02-11 2021-09-17 霍夫曼-拉罗奇有限公司 R-spondin translocations and methods of use thereof
KR102008190B1 (en) 2012-02-15 2019-08-07 바이오버라티브 테라퓨틱스 인크. Recombinant factor viii proteins
ES2935489T3 (en) 2012-02-15 2023-03-07 Bioverativ Therapeutics Inc Factor VIII compositions and methods of preparation and use thereof
WO2013120929A1 (en) 2012-02-15 2013-08-22 F. Hoffmann-La Roche Ag Fc-receptor based affinity chromatography
AU2013240261A1 (en) 2012-03-27 2014-09-18 Genentech, Inc. Diagnosis and treatments relating to HER3 inhibitors
UA117097C2 (en) 2012-03-28 2018-06-25 Санофі Antibodies to bradykinin b1 receptor ligands
AR090549A1 (en) 2012-03-30 2014-11-19 Genentech Inc ANTI-LGR5 AND IMMUNOCATE PLAYERS
CA2867588A1 (en) 2012-03-30 2013-10-03 Genentech, Inc. Diagnostic methods and compositions for treatment of cancer
EP2834270B1 (en) 2012-04-05 2019-10-30 AC Immune S.A. Humanized tau antibody
WO2013163297A1 (en) 2012-04-25 2013-10-31 Momenta Pharmaceuticals, Inc. Modified glycoproteins
WO2013165940A1 (en) 2012-05-01 2013-11-07 Genentech, Inc. Anti-pmel17 antibodies and immunoconjugates
CA2872856A1 (en) 2012-05-07 2013-11-14 Sanofi Methods for preventing biofilm formation
WO2013167153A1 (en) 2012-05-09 2013-11-14 Ganymed Pharmaceuticals Ag Antibodies useful in cancer diagnosis
WO2013170191A1 (en) 2012-05-11 2013-11-14 Genentech, Inc. Methods of using antagonists of nad biosynthesis from nicotinamide
EP2852840B1 (en) 2012-05-23 2019-10-09 F.Hoffmann-La Roche Ag Selection method for therapeutic agents
WO2013175276A1 (en) 2012-05-23 2013-11-28 Argen-X B.V Il-6 binding molecules
WO2013184514A1 (en) 2012-06-04 2013-12-12 Irm Llc Site-specific labeling methods and molecules produced thereby
AU2013270682A1 (en) 2012-06-08 2014-12-11 Biogen Ma Inc. Procoagulant compounds
ES2666126T3 (en) 2012-06-08 2018-05-03 Glenmark Pharmaceuticals S.A. Humanized anti-TrkA antibodies with amino acid substitutions
CN104427995A (en) 2012-06-08 2015-03-18 比奥根艾迪克Ma公司 Chimeric clotting factors
WO2013188855A1 (en) 2012-06-15 2013-12-19 Genentech, Inc. Anti-pcsk9 antibodies, formulations, dosing, and methods of use
US20140051834A1 (en) 2012-06-21 2014-02-20 Hoffmann-La Roche, Inc. Incretin Receptor Ligand Polypeptide Fc-Region Fusion Polypeptides And Conjugates With Altered Fc-Effector Function
US9890215B2 (en) 2012-06-22 2018-02-13 King's College London Vista modulators for diagnosis and treatment of cancer
US9499634B2 (en) 2012-06-25 2016-11-22 Zymeworks Inc. Process and methods for efficient manufacturing of highly pure asymmetric antibodies in mammalian cells
KR20150023889A (en) 2012-06-27 2015-03-05 에프. 호프만-라 로슈 아게 Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof
RU2015100656A (en) 2012-06-27 2016-08-20 Ф. Хоффманн-Ля Рош Аг METHOD FOR PRODUCING ANTIBODY FC-FRAGMENT CONNECTING, INCLUDING AT LEAST ONE CONNECTING GROUP, WHICH SPECIALLY RELATED TO THE TARGET, AND THEIR APPLICATION
AR091649A1 (en) 2012-07-02 2015-02-18 Bristol Myers Squibb Co OPTIMIZATION OF ANTIBODIES THAT FIX THE LYMPHOCYTE ACTIVATION GEN 3 (LAG-3) AND ITS USES
RU2630296C2 (en) 2012-07-04 2017-09-06 Ф. Хоффманн-Ля Рош Аг Antibodies to biotin and application methods
SG11201407420RA (en) 2012-07-04 2015-02-27 Hoffmann La Roche Covalently linked antigen-antibody conjugates
RU2630664C2 (en) 2012-07-04 2017-09-11 Ф. Хоффманн-Ля Рош Аг Theophylline antibodies and methods for their application
CA3188124A1 (en) 2012-07-05 2014-01-09 Genentech, Inc. Expression and secretion system
EP3404105A1 (en) 2012-07-06 2018-11-21 Bioverativ Therapeutics Inc. Cell line expressing single chain factor viii polypeptides and uses thereof
PE20150325A1 (en) 2012-07-09 2015-03-05 Genentech Inc IMMUNOCONJUGATES INCLUDING ANTI-CD22 ANTIBODIES AND DERIVATIVES OF NEMORUBICIN.
AU2013288929A1 (en) 2012-07-09 2014-12-04 Genentech, Inc. Immunoconjugates comprising anti-CD22 antibodies
EA201590172A1 (en) 2012-07-09 2015-09-30 Дженентек, Инк. IMMUNOCONGATES CONTAINING ANTIBODIES TO CD79b
IN2014DN10652A (en) 2012-07-09 2015-09-11 Genentech Inc
JP6603128B2 (en) 2012-07-11 2019-11-06 バイオベラティブ セラピューティクス インコーポレイテッド Complex of factor VIII with XTEN and von Willebrand factor protein and uses thereof
BR112015001459B1 (en) 2012-07-25 2023-02-14 Celldex Therapeutics, Inc ISOLATED ANTIBODY OR FRAGMENT THEREOF, CONJUGATE, USES THEREOF, PHARMACEUTICAL COMPOSITION, POLYNUCLEOTIDE, VECTOR, HOST CELL, ISOLATED CELL, KIT, IN VITRO METHOD TO INHIBIT KIT ACTIVITY, METHOD TO PRODUCE AN ANTIBODY
DK2880170T3 (en) 2012-08-02 2016-10-24 Hoffmann La Roche PROCEDURE FOR PREPARING SOLUBLE FcR AS Fc FUSION WITH INERT IMMUNOGLOBULIN Fc REGION AND APPLICATIONS THEREOF
KR102355959B1 (en) 2012-08-23 2022-01-27 어젠시스 인코포레이티드 Antibody drug conjugates (adc) that bind to 158p1d7 proteins
JOP20200308A1 (en) 2012-09-07 2017-06-16 Novartis Ag IL-18 binding molecules
AU2013312211B2 (en) 2012-09-07 2018-03-29 King's College London VISTA modulators for diagnosis and treatment of cancer
US9790268B2 (en) 2012-09-12 2017-10-17 Genzyme Corporation Fc containing polypeptides with altered glycosylation and reduced effector function
AU2013315499B2 (en) 2012-09-12 2018-08-09 Genzyme Corporation Fc containing polypeptides with altered glycosylation and reduced effector function
CA2887129A1 (en) 2012-10-09 2014-04-17 Igenica, Inc. Anti-c16orf54 antibodies and methods of use thereof
NZ707086A (en) 2012-10-30 2019-07-26 Apexigen Inc Anti-cd40 antibodies and methods of use
EP2914621B1 (en) 2012-11-05 2023-06-07 Foundation Medicine, Inc. Novel ntrk1 fusion molecules and uses thereof
AR093378A1 (en) 2012-11-08 2015-06-03 Hoffmann La Roche BINDING PROTEINS OF ANTIGEN HER3 OF UNION TO THE FORK b OF HER3
EA201892509A1 (en) 2012-11-13 2019-04-30 Дженентек, Инк. ANTIBODIES TO HEMAGGLUTININ AND METHODS OF APPLICATION
EP2733153A1 (en) 2012-11-15 2014-05-21 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for the preparation of immunoconjugates and uses thereof
US9914785B2 (en) 2012-11-28 2018-03-13 Zymeworks Inc. Engineered immunoglobulin heavy chain-light chain pairs and uses thereof
WO2014084859A1 (en) 2012-11-30 2014-06-05 Novartis Ag Molecules and methods for modulating tmem16a activities
PL2928921T3 (en) 2012-12-05 2021-06-28 Novartis Ag Compositions and methods for antibodies targeting epo
EP3686218A1 (en) 2012-12-10 2020-07-29 Biogen MA Inc. Anti-blood dendritic cell antigen 2 antibodies and uses thereof
BR112015014063A2 (en) 2012-12-19 2018-10-30 Amplimmune Inc human anti-b7-h4 antibodies and their uses.
EP2935332B1 (en) 2012-12-21 2021-11-10 MedImmune, LLC Anti-h7cr antibodies
WO2014096015A1 (en) 2012-12-21 2014-06-26 F. Hoffmann-La Roche Ag Disulfide-linked multivalent mhc class i comprising multi-function proteins
AU2013371957B2 (en) 2013-01-02 2018-10-25 Ichnos Sciences SA Antibodies that bind to TL1A and their uses
CA2898326C (en) 2013-01-18 2022-05-17 Foundation Medicine, Inc. Methods of treating cholangiocarcinoma
WO2014116749A1 (en) 2013-01-23 2014-07-31 Genentech, Inc. Anti-hcv antibodies and methods of using thereof
WO2014124006A1 (en) 2013-02-05 2014-08-14 The Johns Hopkins University Nanoparticles for magnetic resonance imaging tracking and methods of making and using thereof
JP2016507523A (en) 2013-02-05 2016-03-10 エンクマフ アーゲー Bispecific antibodies against CD3ε and BCMA
EP2762496A1 (en) 2013-02-05 2014-08-06 EngMab AG Method for the selection of antibodies against BCMA
MX371052B (en) 2013-02-08 2020-01-14 Novartis Ag Anti-il-17a antibodies and their use in treating autoimmune and inflammatory disorders.
CN115925957A (en) 2013-02-08 2023-04-07 Irm责任有限公司 Specific sites for modifying antibodies to make immunoconjugates
WO2014124258A2 (en) 2013-02-08 2014-08-14 Irm Llc Specific sites for modifying antibodies to make immunoconjugates
US10065987B2 (en) 2013-02-12 2018-09-04 Bristol-Myers Squibb Company High pH protein refolding methods
ES2813501T3 (en) 2013-02-12 2021-03-24 Bristol Myers Squibb Co Protein Refolding Methods Based on Tangential Flow Filtration
EP4223772A3 (en) 2013-02-15 2023-10-18 Bioverativ Therapeutics Inc. Optimized factor viii gene
CA2900097A1 (en) 2013-02-22 2014-08-28 F. Hoffmann-La Roche Ag Methods of treating cancer and preventing drug resistance
AR094873A1 (en) 2013-02-25 2015-09-02 Genentech Inc METHODS AND COMPOSITIONS TO DETECT AND TREAT MUTANTS OF AKT RESISTANT TO DRUGS
US20140242083A1 (en) 2013-02-26 2014-08-28 Roche Glycart Ag Anti-mcsp antibodies
US9487587B2 (en) 2013-03-05 2016-11-08 Macrogenics, Inc. Bispecific molecules that are immunoreactive with immune effector cells of a companion animal that express an activating receptor and cells that express B7-H3 and uses thereof
EP2964260A2 (en) 2013-03-06 2016-01-13 F. Hoffmann-La Roche AG Methods of treating and preventing cancer drug resistance
KR102420934B1 (en) 2013-03-11 2022-07-15 젠자임 코포레이션 Hyperglycosylated binding polypeptides
US9498532B2 (en) 2013-03-13 2016-11-22 Novartis Ag Antibody drug conjugates
AP2015008740A0 (en) 2013-03-14 2015-09-30 Macrogenics Inc Bispecific molecules that are immunoreactive with immune effector cells that express an activating receptor and an antigen expressed by a cell infected by a virus and uses thereof
EP3611189A1 (en) 2013-03-14 2020-02-19 Novartis AG Antibodies against notch 3
BR112015022576A2 (en) 2013-03-14 2017-10-24 Genentech Inc pharmaceutical product and its use, kit and method for treating hyperproliferative dysfunction
CN105307683A (en) 2013-03-14 2016-02-03 基因泰克公司 Methods of treating cancer and preventing cancer drug resistance
US9562099B2 (en) 2013-03-14 2017-02-07 Genentech, Inc. Anti-B7-H4 antibodies and immunoconjugates
JP6436965B2 (en) 2013-03-14 2018-12-12 ジェネンテック， インコーポレイテッド Anti-B7-H4 antibody and immunoconjugate
WO2014143739A2 (en) 2013-03-15 2014-09-18 Biogen Idec Ma Inc. Anti-alpha v beta 6 antibodies and uses thereof
US10035859B2 (en) 2013-03-15 2018-07-31 Biogen Ma Inc. Anti-alpha V beta 6 antibodies and uses thereof
KR20150128707A (en) 2013-03-15 2015-11-18 제넨테크, 인크. Compositions and methods for diagnosis and treatment of hepatic cancers
BR112015023752B1 (en) 2013-03-15 2023-11-14 Zyngenia, Inc. MODULAR RECOGNITION DOMAIN (MRD), COMPLEX COMPRISING MRD AND CETUXIMAB, USES OF THE COMPLEX TO INHIBIT ANGIOGENESIS AND TREAT CANCER AND PHARMACEUTICAL COMPOSITION COMPRISING SAID COMPLEX
CA2903772A1 (en) 2013-03-15 2014-09-25 Novartis Ag Antibody drug conjugates
SG11201505926VA (en) 2013-03-15 2015-09-29 Biogen Ma Inc Factor ix polypeptide formulations
BR112015023203A8 (en) 2013-03-15 2018-01-23 Constellation Pharmaceuticals Inc methods for treating cancer, method for increasing the efficiency of cancer treatment, method for delaying and / or preventing cancer development, method for treating an individual with cancer, method for increasing sensitivity for a cancer therapy agent, method for extending a sensitivity period and method for extending the duration of response to cancer therapy.
BR112015023120A2 (en) 2013-03-15 2017-11-21 Genentech Inc method for identifying an individual with a disease or dysfunction, method for predicting the responsiveness of an individual with a disease or dysfunction, method for determining the likelihood that an individual with a disease or dysfunction will exhibit benefit from treatment, method for selecting a therapy, Uses of a pd-11 Axis Binding Antagonist, Assay to Identify an Individual with a Disease, Diagnostic Kit, Method to Evaluate a Treatment Response, and Method to Monitor the Response of a Treated Individual
BR112015021576A2 (en) 2013-03-15 2017-10-10 Dana Farber Cancer Inst Inc therapeutic peptides
PE20151673A1 (en) 2013-03-15 2015-11-27 Genentech Inc IL-22 POLYPEPTIDES AND IL-22 Fc FUSION PROTEINS AND METHODS OF USE
US9598485B2 (en) 2013-03-15 2017-03-21 Ac Immune S.A. Anti-tau antibodies and methods of use
WO2014151680A1 (en) 2013-03-15 2014-09-25 Biogen Idec Ma Inc. Treatment and prevention of acute kidney injury using anti-alpha v beta 5 antibodies
CN105143265A (en) 2013-03-15 2015-12-09 豪夫迈·罗氏有限公司 Anti-crth2 antibodies and their use
RU2019108429A (en) 2013-04-29 2019-05-06 Ф. Хоффманн-Ля Рош Аг MODIFIED ASYMMETRIC ANTIBODIES CONNECTING FC-RECEPTOR AND METHODS OF THEIR APPLICATION
AU2014261630B2 (en) 2013-04-29 2019-05-09 F. Hoffmann-La Roche Ag Human FcRn-binding modified antibodies and methods of use
CN105164158A (en) 2013-04-29 2015-12-16 豪夫迈·罗氏有限公司 Fcrn-binding abolished ANTI-IGF-1R antibodies and their use in the treatment of vascular eye diseases
JP6612214B2 (en) 2013-05-20 2019-11-27 ジェネンテック， インコーポレイテッド Anti-transferrin receptor antibodies and methods of use
CN106414489A (en) 2013-05-24 2017-02-15 米迪缪尼有限公司 Anti-b7-h5 antibodies and their uses
CA2922704A1 (en) 2013-05-24 2014-11-27 Neil R. Cashman Cell senescence markers as diagnostic and therapeutic targets
RU2655439C2 (en) 2013-05-31 2018-05-28 Займворкс Инк. Heteromultimers with reduced or silenced effector function
JP2016521715A (en) 2013-06-14 2016-07-25 バイエル ファーマ アクチエンゲゼルシャフト Anti-TWEAKR antibody and use thereof
AR096601A1 (en) 2013-06-21 2016-01-20 Novartis Ag ANTIBODIES OF LEXINED OXIDATED LDL RECEIVER 1 AND METHODS OF USE
US9562101B2 (en) 2013-06-21 2017-02-07 Novartis Ag Lectin-like oxidized LDL receptor 1 antibodies and methods of use
EP3022224A2 (en) 2013-07-18 2016-05-25 Fabrus, Inc. Antibodies with ultralong complementarity determining regions
AU2014290361B2 (en) 2013-07-18 2019-04-18 Taurus Biosciences, Llc Humanized antibodies with ultralong complementarity determining regions
WO2015014376A1 (en) 2013-07-31 2015-02-05 Biontech Ag Diagnosis and therapy of cancer involving cancer stem cells
RU2679657C2 (en) 2013-08-01 2019-02-12 Эдженсис, Инк. Antibody drug conjugates that bind to cd37 proteins
LT3708583T (en) 2013-08-01 2022-04-11 Five Prime Therapeutics, Inc. Afucosylated anti-fgfr2iiib antibodies
EP3875106A1 (en) 2013-08-08 2021-09-08 Bioverativ Therapeutics Inc. Purification of chimeric fviii molecules
UA116479C2 (en) 2013-08-09 2018-03-26 Макродженікс, Інк. Bi-specific monovalent fc diabodies that are capable of binding cd32b and cd79b and uses thereof
US11384149B2 (en) 2013-08-09 2022-07-12 Macrogenics, Inc. Bi-specific monovalent Fc diabodies that are capable of binding CD32B and CD79b and uses thereof
EA201690213A1 (en) 2013-08-12 2016-07-29 Дженентек, Инк. COMPOSITIONS AND METHOD FOR TREATMENT OF CONNECTED STATES
TWI592426B (en) 2013-08-13 2017-07-21 賽諾菲公司 Antibodies to plasminogen activator inhibitor-1 (pai-1) and uses thereof
US9845363B2 (en) 2013-08-13 2017-12-19 Sanofi Antibodies to plasminogen activator inhibitor-1 (PAI-1) and uses thereof
MX2016001969A (en) 2013-08-14 2016-06-02 Novartis Ag Methods of treating sporadic inclusion body myositis.
US10548953B2 (en) 2013-08-14 2020-02-04 Bioverativ Therapeutics Inc. Factor VIII-XTEN fusions and uses thereof
PL3036320T3 (en) 2013-08-19 2021-11-02 Biogen Ma Inc. Control of protein glycosylation by culture medium supplementation and cell culture process parameters
US20150056199A1 (en) 2013-08-22 2015-02-26 Acceleron Pharma, Inc. Tgf-beta receptor type ii variants and uses thereof
EP2840091A1 (en) 2013-08-23 2015-02-25 MacroGenics, Inc. Bi-specific diabodies that are capable of binding gpA33 and CD3 and uses thereof
EP2839842A1 (en) 2013-08-23 2015-02-25 MacroGenics, Inc. Bi-specific monovalent diabodies that are capable of binding CD123 and CD3 and uses thereof
HUE049281T2 (en) 2013-09-13 2020-09-28 Beigene Switzerland Gmbh Anti-pd1 antibodies and their use as therapeutics and diagnostics
MX2016003248A (en) 2013-09-17 2016-06-07 Genentech Inc Methods of using anti-lgr5 antibodies.
CN104436157A (en) 2013-09-23 2015-03-25 恩金生物有限公司 Influenza vaccine and therapy
EP3903599A1 (en) 2013-09-25 2021-11-03 Bioverativ Therapeutics Inc. On-column viral inactivation methods
WO2015044083A1 (en) 2013-09-27 2015-04-02 F. Hoffmann-La Roche Ag Thermus thermophilus slyd fkbp domain specific antibodies
WO2015050959A1 (en) 2013-10-01 2015-04-09 Yale University Anti-kit antibodies and methods of use thereof
BR112016006999B1 (en) 2013-10-02 2023-11-14 Medimmune, Llc ANTI-INFLUENZA NEUTRALIZING ANTIBODIES AND THEIR USES, THEIR PRODUCTION METHOD, COMPOSITION THAT COMPRISES THEM, NUCLEIC ACID AND VECTOR
MX2016003593A (en) 2013-10-11 2016-06-02 Hoffmann La Roche Multispecific domain exchanged common variable light chain antibodies.
BR112016007635A2 (en) 2013-10-11 2017-09-12 Genentech Inc nsp4 inhibitors and methods of use
CA2925598A1 (en) 2013-10-18 2015-04-23 Genentech, Inc. Anti-rspo antibodies and methods of use
WO2015057939A1 (en) 2013-10-18 2015-04-23 Biogen Idec Ma Inc. Anti-s1p4 antibodies and uses thereof
MX2016005159A (en) 2013-10-23 2016-07-05 Genentech Inc Methods of diagnosing and treating eosinophilic disorders.
KR102354787B1 (en) 2013-10-25 2022-01-24 악셀레론 파마 인코포레이티드 Endoglin peptides to treat fibrotic diseases
ES2759252T3 (en) 2013-10-31 2020-05-08 Resolve Therapeutics Llc Nuclease-albumin fusions and therapeutic methods
ES2865196T3 (en) 2013-11-07 2021-10-15 Inst Nat Sante Rech Med Neuregulin allosteric anti-human HER3 antibodies
CN104623637A (en) 2013-11-07 2015-05-20 健能隆医药技术（上海）有限公司 Application of IL-22 dimer in preparation of intravenous injection drugs
EP3065769A4 (en) 2013-11-08 2017-05-31 Biogen MA Inc. Procoagulant fusion compound
SI3071597T1 (en) 2013-11-21 2020-11-30 F. Hoffmann-La Roche Ag Anti-alpha-synuclein antibodies and methods of use
CA2931356A1 (en) 2013-11-27 2015-06-04 Zymeworks Inc. Bispecific antigen-binding constructs targeting her2
EP3077504B1 (en) 2013-12-06 2019-08-14 Dana-Farber Cancer Institute, Inc. Therapeutic peptides
CN114044825A (en) 2013-12-09 2022-02-15 爱乐科斯公司 anti-Siglec-8 antibodies and methods of use thereof
CA2931340A1 (en) 2013-12-13 2015-06-18 Genentech, Inc. Anti-cd33 antibodies and immunoconjugates
US8986691B1 (en) 2014-07-15 2015-03-24 Kymab Limited Method of treating atopic dermatitis or asthma using antibody to IL4RA
US20150190506A1 (en) 2013-12-17 2015-07-09 Genentech, Inc. Combination therapy comprising ox40 binding agonists and pd-1 axis binding antagonists
PE20210107A1 (en) 2013-12-17 2021-01-19 Genentech Inc ANTI-CD3 ANTIBODIES AND METHODS OF USE
US20150210772A1 (en) 2013-12-17 2015-07-30 Genentech, Inc. Methods of treating cancer using pd-1 axis binding antagonists and an anti-cd20 antibody
AU2014364601A1 (en) 2013-12-17 2016-07-07 Genentech, Inc. Methods of treating HER2-positive cancers using PD-1 axis binding antagonists and anti-HER2 antibodies
US8980273B1 (en) 2014-07-15 2015-03-17 Kymab Limited Method of treating atopic dermatitis or asthma using antibody to IL4RA
AU2014368696A1 (en) 2013-12-20 2016-06-02 F. Hoffmann-La Roche Ag Humanized anti-Tau(pS422) antibodies and methods of use
TWI728373B (en) 2013-12-23 2021-05-21 美商建南德克公司 Antibodies and methods of use
EP3087098B1 (en) 2013-12-24 2020-04-08 Janssen Pharmaceutica NV Anti-vista antibodies and fragments
CN112142843A (en) 2013-12-24 2020-12-29 阿尔金克斯有限公司 FcRn antagonists and methods of use
US11014987B2 (en) 2013-12-24 2021-05-25 Janssen Pharmaceutics Nv Anti-vista antibodies and fragments, uses thereof, and methods of identifying same
KR20160104628A (en) 2014-01-03 2016-09-05 에프. 호프만-라 로슈 아게 Bispecific anti-hapten/anti-blood brain barrier receptor antibodies, complexes thereof and their use as blood brain barrier shuttles
KR102278429B1 (en) 2014-01-03 2021-07-16 에프. 호프만-라 로슈 아게 Covalently linked polypeptide toxin-antibody conjugates
CN105873615B (en) 2014-01-03 2020-12-25 豪夫迈·罗氏有限公司 Covalently linked helicar-anti-helicar antibody conjugates and uses thereof
CA2932547C (en) 2014-01-06 2023-05-23 F. Hoffmann-La Roche Ag Monovalent blood brain barrier shuttle modules
SG11201605242YA (en) 2014-01-10 2016-07-28 Biogen Ma Inc Factor viii chimeric proteins and uses thereof
KR20160107190A (en) 2014-01-15 2016-09-13 에프. 호프만-라 로슈 아게 Fc-region variants with modified fcrn- and maintained protein a-binding properties
KR20160111469A (en) 2014-01-24 2016-09-26 제넨테크, 인크. Methods of using anti-steap1 antibodies and immunoconjugates
DK3097122T3 (en) 2014-01-24 2020-08-10 Ngm Biopharmaceuticals Inc ANTIBODIES BINDING BETA-KLOTHO DOMAIN 2 AND METHODS OF USING IT
CA2937539A1 (en) 2014-02-04 2015-08-13 Genentech, Inc. Mutant smoothened and methods of using the same
TWI705824B (en) 2014-02-08 2020-10-01 美商建南德克公司 Methods of treating alzheimer's disease
ES2873248T3 (en) 2014-02-08 2021-11-03 Hoffmann La Roche Methods to treat Alzheimer's disease
SG10201808259TA (en) 2014-02-12 2018-10-30 Genentech Inc Anti-jagged1 antibodies and methods of use
EP3107574A2 (en) 2014-02-21 2016-12-28 F. Hoffmann-La Roche AG Anti-il-13/il-17 bispecific antibodies and uses thereof
DK3110446T3 (en) 2014-02-28 2022-02-28 Allakos Inc Methods and compositions for treating Siglec-8-associated diseases
WO2015134671A1 (en) 2014-03-04 2015-09-11 Targeson, Inc. Molecular imaging contrast agents and uses thereof
NZ631007A (en) 2014-03-07 2015-10-30 Alexion Pharma Inc Anti-c5 antibodies having improved pharmacokinetics
AU2015229463A1 (en) 2014-03-12 2016-09-15 Novartis Ag Specific sites for modifying antibodies to make immunoconjugates
CN106103730B (en) 2014-03-14 2021-06-08 豪夫迈·罗氏有限公司 Methods and compositions for secretion of heterologous polypeptides
CN113491766A (en) 2014-03-14 2021-10-12 达纳－法伯癌症研究所公司 Vaccine compositions and methods for restoring function of the NKG2D pathway against cancer
SI3129067T1 (en) 2014-03-19 2023-06-30 Genzyme Corporation Site-specific glycoengineering of targeting moieties
CN106164094B (en) 2014-03-21 2021-05-14 X博迪公司 Bispecific antigen binding polypeptides
WO2015140591A1 (en) 2014-03-21 2015-09-24 Nordlandssykehuset Hf Anti-cd14 antibodies and uses thereof
JP2017516458A (en) 2014-03-24 2017-06-22 ジェネンテック， インコーポレイテッド Cancer treatment with c-met antagonist and correlation with HGF expression of c-met antagonist
EP3126386A1 (en) 2014-03-31 2017-02-08 F. Hoffmann-La Roche AG Combination therapy comprising anti-angiogenesis agents and ox40 binding agonists
KR20160145624A (en) 2014-03-31 2016-12-20 제넨테크, 인크. Anti-ox40 antibodies and methods of use
JP6614582B2 (en) 2014-04-04 2019-12-04 バイオノミクス インコーポレイテッド Humanized antibody that binds to LGR5
EP3145530B1 (en) 2014-04-21 2021-04-07 D&D Pharmatech Inc. Trail receptor agonists for treatment of fibrotic diseases
TW201622746A (en) 2014-04-24 2016-07-01 諾華公司 Methods of improving or accelerating physical recovery after surgery for hip fracture
WO2015164615A1 (en) 2014-04-24 2015-10-29 University Of Oslo Anti-gluten antibodies and uses thereof
ES2883837T3 (en) 2014-05-02 2021-12-09 The Res Institute At Nationwide Childrens Hospital Compositions and methods for immunomodulation of anti-LYST
RU2016150370A (en) 2014-05-22 2018-06-26 Дженентек, Инк. Antibodies and immunoconjugates against GPC3
WO2015179835A2 (en) 2014-05-23 2015-11-26 Genentech, Inc. Mit biomarkers and methods using the same
US10155818B2 (en) 2014-05-28 2018-12-18 Agenus Inc. Anti-GITR antibodies and methods of use thereof
SG10201913680PA (en) 2014-05-29 2020-03-30 Macrogenics Inc Tri-specific binding molecules that specifically bind to multiple cancer antigens and methods of use thereof
ES2905777T3 (en) 2014-05-30 2022-04-12 Shanghai Henlius Biotech Inc Anti-epidermal growth factor receptor (EGFR) antibodies
PE20170441A1 (en) 2014-06-06 2017-04-26 Bristol Myers Squibb Co ANTIBODIES AGAINST THE GLUCOCORTICOID-INDUCED TUMOR NECROSIS FACTOR RECEPTOR (GITR) AND ITS USES
CN107073109B (en) 2014-06-11 2021-08-06 凯西·A·格林 Use of VISTA agonists and antagonists to inhibit or enhance humoral immunity
EP3155015A1 (en) 2014-06-11 2017-04-19 F. Hoffmann-La Roche AG Anti-lgr5 antibodies and uses thereof
US20230190750A1 (en) 2014-06-13 2023-06-22 Genentech, Inc. Methods of treating and preventing cancer drug resistance
US20170291939A1 (en) 2014-06-25 2017-10-12 Novartis Ag Antibodies specific for il-17a fused to hyaluronan binding peptide tags
EP3164419A1 (en) 2014-06-26 2017-05-10 F. Hoffmann-La Roche AG Anti-brdu antibodies and methods of use
AR100978A1 (en) 2014-06-26 2016-11-16 Hoffmann La Roche ANTI-Tau HUMANIZED ANTIBODY BRAIN LAUNCHERS (pS422) AND USES OF THE SAME
US11008561B2 (en) 2014-06-30 2021-05-18 Bioverativ Therapeutics Inc. Optimized factor IX gene
WO2016000619A1 (en) 2014-07-03 2016-01-07 Beigene, Ltd. Anti-pd-l1 antibodies and their use as therapeutics and diagnostics
RU2017103289A (en) 2014-07-11 2018-08-14 Дженентек, Инк. INHIBITING THE NOTCH WAY
EP3166974A1 (en) 2014-07-11 2017-05-17 Genentech, Inc. Anti-pd-l1 antibodies and diagnostic uses thereof
US10294292B2 (en) 2014-07-15 2019-05-21 Medimmune, Llc Neutralizing anti-influenza B antibodies and uses thereof
RU2711141C2 (en) 2014-07-22 2020-01-15 СиБи ТЕРЕПЬЮТИКС, ИНК. Anti-pd-1 antibodies
WO2016019126A1 (en) 2014-07-30 2016-02-04 The Research Foundation For The State University Of New York System and method for delivering genetic material or protein to cells
EP3174906A1 (en) 2014-08-01 2017-06-07 Massachusetts Institute Of Technology Modified alginates for anti-fibrotic materials and applications
NZ728749A (en) 2014-08-05 2023-06-30 Cb Therapeutics Inc Anti-pd-l1 antibodies
WO2016020791A1 (en) 2014-08-05 2016-02-11 Novartis Ag Ckit antibody drug conjugates
EP3194437B1 (en) 2014-08-07 2021-01-20 Novartis AG Angiopoietin-like 4 (angptl4) antibodies and methods of use
CU24453B1 (en) 2014-08-07 2019-11-04 Novartis Ag ANTI-PROTEIN ANTIBODIES SIMILAR TO ANGIOPOYETIN 4
US10238748B2 (en) 2014-08-12 2019-03-26 Novartis Ag Anti-CDH6 antibody drug conjugates
UA122395C2 (en) 2014-08-19 2020-11-10 Мерк Шарп Енд Доум Корп. ANTIBODY AGAINST TIGIT
JO3663B1 (en) 2014-08-19 2020-08-27 Merck Sharp & Dohme Anti-lag3 antibodies and antigen-binding fragments
RU2764074C2 (en) 2014-08-28 2022-01-13 Байоатла, Ллк Conditionally active chimeric antigen receptors for modified t-cells
JP2017528468A (en) 2014-09-10 2017-09-28 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ Galactose engineered immunoglobulin 1 antibody
CN113698488A (en) 2014-09-12 2021-11-26 基因泰克公司 anti-B7-H4 antibodies and immunoconjugates
AU2015314826A1 (en) 2014-09-12 2017-03-02 Genentech, Inc. Cysteine engineered antibodies and conjugates
AU2015314744A1 (en) 2014-09-12 2017-03-02 Genentech, Inc. Anti-CLL-1 antibodies and immunoconjugates
MA40576B1 (en) 2014-09-12 2020-11-30 Genentech Inc Anti-her2 antibodies and immunoconjugates
EP3197500A1 (en) 2014-09-17 2017-08-02 Genentech, Inc. Immunoconjugates comprising anti-her2 antibodies and pyrrolobenzodiazepines
PL3262071T3 (en) 2014-09-23 2020-08-10 F. Hoffmann-La Roche Ag Method of using anti-cd79b immunoconjugates
SG11201701803XA (en) 2014-09-26 2017-04-27 Bayer Pharma AG Stabilized adrenomedullin derivatives and use thereof
BR112017006515A8 (en) 2014-09-29 2018-02-27 Univ Duke bispecific molecules, compositions, method for treating or preventing HIV-1 infection in an individual with this need, and vector
MA41044A (en) 2014-10-08 2017-08-15 Novartis Ag COMPOSITIONS AND METHODS OF USE FOR INCREASED IMMUNE RESPONSE AND CANCER TREATMENT
AU2015330869B2 (en) 2014-10-09 2021-07-08 Genzyme Corporation Glycoengineered antibody drug conjugates
ES2753391T3 (en) 2014-10-14 2020-04-08 Halozyme Inc Adenosine deaminase 2 (ADA2) compositions, variants thereof and methods of use thereof
US9732148B2 (en) 2014-10-16 2017-08-15 Genentech, Inc. Anti-α-synuclein antibodies and methods of use
MA41480A (en) 2014-10-17 2017-12-19 Glenmark Pharmaceuticals Sa ANTIBODIES BOUND TO CCR6 AND THEIR USES
MA41685A (en) 2014-10-17 2017-08-22 Biogen Ma Inc COPPER SUPPLEMENT FOR THE REGULATION OF GLYCOSYLATION IN A MAMMAL CELL CULTURE PROCESS
WO2016070001A1 (en) 2014-10-31 2016-05-06 Jounce Therapeutics, Inc. Methods of treating conditions with antibodies that bind b7-h4
US20160160290A1 (en) 2014-11-03 2016-06-09 Genentech, Inc. Methods and biomarkers for predicting efficacy and evaluation of an ox40 agonist treatment
CA2966523A1 (en) 2014-11-03 2016-05-12 Genentech, Inc. Assays for detecting t cell immune subsets and methods of use thereof
RU2739500C2 (en) 2014-11-05 2020-12-25 Дженентек, Инк. Methods for producing double-stranded proteins in bacteria
BR112017008666A2 (en) 2014-11-05 2018-01-30 Genentech, Inc. anti-fgfr2 / 3 antibodies and methods of use
AU2015342961B2 (en) 2014-11-05 2021-08-12 Genentech, Inc. Methods of producing two chain proteins in bacteria
SG11201703376QA (en) 2014-11-06 2017-05-30 Genentech Inc Combination therapy comprising ox40 binding agonists and tigit inhibitors
CN107074966A (en) 2014-11-06 2017-08-18 豪夫迈·罗氏有限公司 The Fc region variants of FCRN and albumin A binding property with change
EP3215528B1 (en) 2014-11-06 2019-08-07 F.Hoffmann-La Roche Ag Fc-region variants with modified fcrn-binding and methods of use
WO2016073157A1 (en) 2014-11-06 2016-05-12 Genentech, Inc. Anti-ang2 antibodies and methods of use thereof
CN107105632A (en) 2014-11-10 2017-08-29 豪夫迈·罗氏有限公司 Nephrosis animal model and its therapeutic agent
CN107172879B (en) 2014-11-10 2021-11-05 豪夫迈·罗氏有限公司 Anti-interleukin-33 antibodies and uses thereof
CA2967188A1 (en) 2014-11-14 2016-05-19 Novartis Ag Antibody drug conjugates
CN106999583A (en) 2014-11-17 2017-08-01 豪夫迈·罗氏有限公司 Combination treatment comprising OX40 combinations activator and the axle binding antagonists of PD 1
HUE053239T2 (en) 2014-11-19 2021-06-28 Axon Neuroscience Se Humanized tau antibodies in alzheimer's disease
EP3221361B1 (en) 2014-11-19 2021-04-21 Genentech, Inc. Anti-transferrin receptor / anti-bace1 multispecific antibodies and methods of use
JP6779876B2 (en) 2014-11-19 2020-11-04 ジェネンテック， インコーポレイテッド Anti-transferrin receptor antibody and how to use it
EP3845565A3 (en) 2014-11-19 2021-09-08 Genentech, Inc. Antibodies against bace1 and use thereof for neural disease immunotherapy
CN113773388A (en) 2014-11-21 2021-12-10 百时美施贵宝公司 anti-CD 73 antibodies and uses thereof
MX2017006323A (en) 2014-11-21 2017-08-21 Bristol Myers Squibb Co Antibodies comprising modified heavy constant regions.
WO2016087416A1 (en) 2014-12-03 2016-06-09 F. Hoffmann-La Roche Ag Multispecific antibodies
EP3226900A4 (en) 2014-12-05 2018-09-19 Immunext, Inc. Identification of vsig8 as the putative vista receptor and its use thereof to produce vista/vsig8 modulators
LT3227336T (en) 2014-12-05 2019-09-25 F. Hoffmann-La Roche Ag Anti-cd79b antibodies and methods of use
US10398774B2 (en) 2014-12-09 2019-09-03 INSERM (Institut National de la Santé et de la Recherche Médicale) Human monoclonal antibodies against AXL
KR20170085595A (en) 2014-12-10 2017-07-24 제넨테크, 인크. Blood brain barrier receptor antibodies and methods of use
AU2015365167B2 (en) 2014-12-19 2021-07-29 Chugai Seiyaku Kabushiki Kaisha Anti-C5 antibodies and methods of use
UY36449A (en) 2014-12-19 2016-07-29 Novartis Ag COMPOSITIONS AND METHODS FOR ANTIBODIES DIRECTED TO BMP6
CN113563468A (en) 2014-12-19 2021-10-29 雷根尼桑斯公司 Antibodies that bind to human C6 and uses thereof
KR102644115B1 (en) 2014-12-23 2024-03-05 브리스톨-마이어스 스큅 컴퍼니 Antibodies to tigit
WO2016111947A2 (en) 2015-01-05 2016-07-14 Jounce Therapeutics, Inc. Antibodies that inhibit tim-3:lilrb2 interactions and uses thereof
SG10201906471PA (en) 2015-01-14 2019-09-27 Brigham & Womens Hospital Inc Treatment of cancer with anti-lap monoclonal antibodies
JP2018511557A (en) 2015-01-22 2018-04-26 中外製薬株式会社 Combination and use of two or more anti-C5 antibodies
AU2015380455A1 (en) 2015-01-26 2017-08-03 Macrogenics, Inc. Multivalent molecules comprising DR5-binding domains
WO2016123125A1 (en) 2015-01-27 2016-08-04 The Johns Hopkins University Hypotonic hydrogel formulations for enhanced transport of active agents at mucosal surfaces
JP2018506275A (en) 2015-01-28 2018-03-08 ジェネンテック， インコーポレイテッド Gene expression markers and treatment of multiple sclerosis
JP2018512597A (en) 2015-02-04 2018-05-17 ジェネンテック， インコーポレイテッド Mutant smoothened and method of using the same
CN114773469A (en) 2015-02-05 2022-07-22 中外制药株式会社 Antibodies comprising an ion concentration-dependent antigen-binding domain, FC region variants, IL-8-binding antibodies and uses thereof
EP3259597B1 (en) 2015-02-19 2022-04-06 Compugen Ltd. Pvrig polypeptides and methods of treatment
US10227408B2 (en) 2015-02-19 2019-03-12 Compugen Ltd. Anti-PVRIG antibodies and methods of use
WO2016135041A1 (en) 2015-02-26 2016-09-01 INSERM (Institut National de la Santé et de la Recherche Médicale) Fusion proteins and antibodies comprising thereof for promoting apoptosis
US10711067B2 (en) 2015-03-03 2020-07-14 Xoma (Us) Llc Treatment of post-prandial hyperinsulinemia and hypoglycemia after bariatric surgery
MY188761A (en) 2015-03-09 2021-12-29 Argenx Bvba Method of reducing serum levels of fc-containing agents using fcrn antagonists
AU2016233398A1 (en) 2015-03-16 2017-09-07 F. Hoffmann-La Roche Ag Methods of detecting and quantifying IL-13 and uses in diagnosing and treating Th2-associated diseases
WO2016146833A1 (en) 2015-03-19 2016-09-22 F. Hoffmann-La Roche Ag Biomarkers for nad(+)-diphthamide adp ribosyltransferase resistance
PL3273992T3 (en) 2015-03-23 2020-11-16 Jounce Therapeutics, Inc. Antibodies to icos
DK3274370T3 (en) 2015-03-23 2020-02-17 Bayer Pharma AG ANTI-CEACAM6 ANTIBODIES AND APPLICATIONS THEREOF
US10167334B2 (en) 2015-04-03 2019-01-01 Xoma Technology Ltd. Treatment of cancer using anti-TGF-BETA and PD-1 antibodies
CN114456272A (en) 2015-04-03 2022-05-10 优瑞科生物技术公司 Constructs targeting AFP peptide/MHC complexes and uses thereof
CN107709364A (en) 2015-04-07 2018-02-16 豪夫迈·罗氏有限公司 Antigen binding complex and application method with agonist activity
LT3283508T (en) 2015-04-17 2021-07-12 Alpine Immune Sciences, Inc. Immunomodulatory proteins with tunable affinities
JP7044553B2 (en) 2015-04-24 2022-03-30 ジェネンテック， インコーポレイテッド How to identify bacteria containing bound polypeptides
JP2018520642A (en) 2015-05-01 2018-08-02 ジェネンテック， インコーポレイテッド Mask anti-CD3 antibody and method of use thereof
WO2016179194A1 (en) 2015-05-04 2016-11-10 Jounce Therapeutics, Inc. Lilra3 and method of using the same
TWI820377B (en) 2015-05-07 2023-11-01 美商艾吉納斯公司 Anti-ox40 antibodies and methods of use thereof
CN116196414A (en) 2015-05-11 2023-06-02 豪夫迈·罗氏有限公司 Compositions and methods for treating lupus nephritis
IL295002A (en) 2015-05-12 2022-09-01 Genentech Inc Therapeutic and diagnostic methods for cancer comprising a pd-l1 binding antagonist
JP6770533B2 (en) 2015-05-22 2020-10-14 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル A human monoclonal antibody fragment that inhibits both the catalytic activity of Cath-D and its binding to the LRP1 receptor.
HUE061253T2 (en) 2015-05-29 2023-06-28 Bristol Myers Squibb Co Antibodies against ox40 and uses thereof
EP3303619B1 (en) 2015-05-29 2020-06-10 H. Hoffnabb-La Roche Ag Pd-l1 promoter methylation in cancer
WO2016196343A1 (en) 2015-05-29 2016-12-08 Genentech, Inc. Humanized anti-ebola virus glycoprotein antibodies and methods of use
KR20180012753A (en) 2015-05-29 2018-02-06 제넨테크, 인크. Treatment and Diagnosis Methods for Cancer
US20180258143A1 (en) 2015-05-30 2018-09-13 Molecular Templates, Inc. De-Immunized, Shiga Toxin A Subunit Scaffolds and Cell-Targeting Molecules Comprising the Same
MX2017015189A (en) 2015-06-01 2018-04-13 Medimmune Llc Neutralizing anti-influenza binding molecules and uses thereof.
JP2018516933A (en) 2015-06-02 2018-06-28 ジェネンテック， インコーポレイテッド Compositions and methods for treating neurological disorders using anti-IL-34 antibodies
BR112017021484A2 (en) 2015-06-05 2018-07-03 Novartis Ag antibodies targeting bone morphogenetic protein 9 (bmp9) and methods for them
SG10201911349YA (en) 2015-06-05 2020-01-30 Genentech Inc Anti-tau antibodies and methods of use
TWI773646B (en) 2015-06-08 2022-08-11 美商宏觀基因股份有限公司 Lag-3-binding molecules and methods of use thereof
AU2016274585A1 (en) 2015-06-08 2017-12-14 Genentech, Inc. Methods of treating cancer using anti-OX40 antibodies
KR20180025888A (en) 2015-06-08 2018-03-09 제넨테크, 인크. Methods for treating cancer using anti-OX40 antibodies and PD-1 axis-binding antagonists
TW201709929A (en) 2015-06-12 2017-03-16 宏觀基因股份有限公司 Combination therapy for the treatment of cancer
US10017577B2 (en) 2015-06-15 2018-07-10 Genentech, Inc. Antibodies and immunoconjugates
PE20180330A1 (en) 2015-06-16 2018-02-13 Genentech Inc AFFINITY MATURED AND HUMANIZED ANTIBODIES FOR FCRH5 AND METHODS FOR THEIR USE
WO2016204966A1 (en) 2015-06-16 2016-12-22 Genentech, Inc. Anti-cd3 antibodies and methods of use
CN107847568B (en) 2015-06-16 2022-12-20 豪夫迈·罗氏有限公司 anti-CLL-1 antibodies and methods of use
KR20180018538A (en) 2015-06-17 2018-02-21 제넨테크, 인크. Methods for the treatment of locally advanced or metastatic breast cancer using PD-1 axis-binding antagonists and taxanes
JP2018524312A (en) 2015-06-17 2018-08-30 ジェネンテック， インコーポレイテッド Anti-HER2 antibody and method of use
WO2016205567A1 (en) 2015-06-17 2016-12-22 Allakos Inc. Methods and compositions for treating fibrotic diseases
US20190194315A1 (en) 2015-06-17 2019-06-27 Novartis Ag Antibody drug conjugates
EP3313877B1 (en) 2015-06-24 2020-06-03 H. Hoffnabb-La Roche Ag Humanized anti-tau(ps422) antibodies and methods of use
WO2016207717A1 (en) 2015-06-24 2016-12-29 Janssen Pharmaceutica Nv Anti-vista antibodies and fragments
CN113999312A (en) 2015-06-24 2022-02-01 豪夫迈·罗氏有限公司 Anti-transferrin receptor antibodies with tailored affinity
JOP20200312A1 (en) 2015-06-26 2017-06-16 Novartis Ag Factor xi antibodies and methods of use
CN108473573A (en) 2015-06-29 2018-08-31 豪夫迈·罗氏有限公司 II type anti-CD 20 antibodies are used in organ transplant
EP3514174B1 (en) 2015-06-29 2021-03-31 Ventana Medical Systems, Inc. Materials and methods for performing histochemical assays for human pro-epiregulin and amphiregulin
WO2017005847A1 (en) 2015-07-07 2017-01-12 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies having specificity to myosin 18a and uses thereof
WO2017009712A1 (en) 2015-07-13 2017-01-19 Compugen Ltd. Hide1 compositions and methods
IL256879B1 (en) * 2015-07-24 2024-01-01 Gliknik Inc Fusion proteins of human protein fragments to create orderly multimerized immunoglobulin fc compositions with enhanced complement binding
KR20180093127A (en) 2015-07-30 2018-08-20 마크로제닉스, 인크. PD-1-Binding Molecules and Methods of Use Thereof
EP3670535A1 (en) 2015-08-03 2020-06-24 EngMab Sàrl Monoclonal antibodies against bcma
CN108350072B (en) 2015-08-03 2022-05-24 诺华股份有限公司 Methods of treating FGF 21-associated disorders
EA201890423A1 (en) 2015-08-03 2018-07-31 Биовератив Терапьютикс Инк. SLIGHT PROTEINS OF THE FACTOR IX, METHODS OF THEIR RECEPTION AND APPLICATION
EP4218792A1 (en) 2015-08-04 2023-08-02 Acceleron Pharma Inc. Composition for treating myeloproliferative disorders
WO2017024285A2 (en) 2015-08-06 2017-02-09 Xoma (Us) Llc Antibody fragments against the insulin receptor and uses thereof to treat hypoglycemia
CN105384825B (en) 2015-08-11 2018-06-01 南京传奇生物科技有限公司 A kind of bispecific chimeric antigen receptor and its application based on single domain antibody
CN108026180B (en) 2015-08-28 2022-06-07 豪夫迈·罗氏有限公司 Anti-hypusine antibodies and uses thereof
PE20181322A1 (en) 2015-09-01 2018-08-14 Agenus Inc ANTI-PD1 ANTIBODY AND ITS METHODS OF USE
TN2018000076A1 (en) 2015-09-09 2019-07-08 Novartis Ag Thymic stromal lymphopoietin (tslp)-binding molecules and methods of using the molecules
HUE053914T2 (en) 2015-09-09 2021-07-28 Novartis Ag Thymic stromal lymphopoietin (tslp)-binding antibodies and methods of using the antibodies
US20190022092A1 (en) 2015-09-15 2019-01-24 Acerta Pharma B.V. Therapeutic Combinations of a BTK Inhibitor and a GITR Binding Molecule, a 4-1BB Agonist, or an OX40 Agonist
US9862760B2 (en) 2015-09-16 2018-01-09 Novartis Ag Polyomavirus neutralizing antibodies
CA2999369C (en) 2015-09-22 2023-11-07 Spring Bioscience Corporation Anti-ox40 antibodies and diagnostic uses thereof
AU2016326666B2 (en) 2015-09-23 2023-06-15 Genentech, Inc. Optimized variants of anti-VEGF antibodies
CN108289954B (en) 2015-09-24 2022-05-31 阿布维特罗有限责任公司 HIV antibody compositions and methods of use
WO2017058859A1 (en) 2015-09-29 2017-04-06 Celgene Corporation Pd-1 binding proteins and methods of use thereof
AR106188A1 (en) 2015-10-01 2017-12-20 Hoffmann La Roche ANTI-CD19 HUMANIZED HUMAN ANTIBODIES AND METHODS OF USE
AR106189A1 (en) 2015-10-02 2017-12-20 Hoffmann La Roche BIESPECTIFIC ANTIBODIES AGAINST HUMAN A-b AND THE HUMAN TRANSFERRINE RECEIVER AND METHODS OF USE
MA43345A (en) 2015-10-02 2018-08-08 Hoffmann La Roche PYRROLOBENZODIAZEPINE ANTIBODY-DRUG CONJUGATES AND METHODS OF USE
AU2016333512B2 (en) 2015-10-02 2022-11-17 F. Hoffmann-La Roche Ag Bispecific anti-human CD20/human transferrin receptor antibodies and methods of use
EP3150636A1 (en) 2015-10-02 2017-04-05 F. Hoffmann-La Roche AG Tetravalent multispecific antibodies
US11891440B2 (en) * 2015-10-05 2024-02-06 Circle33 Llc Antibodies with improved stability to intestinal digestion, polynucleotides thereof and methods of use thereof to treat disease
JP2018529747A (en) 2015-10-06 2018-10-11 ジェネンテック， インコーポレイテッド Methods for treating multiple sclerosis
WO2017066714A1 (en) 2015-10-16 2017-04-20 Compugen Ltd. Anti-vsig1 antibodies and drug conjugates
MA43354A (en) 2015-10-16 2018-08-22 Genentech Inc CONJUGATE DRUG CONJUGATES WITH CLOUDY DISULPHIDE
MA45326A (en) 2015-10-20 2018-08-29 Genentech Inc CALICHEAMICIN-ANTIBODY-DRUG CONJUGATES AND METHODS OF USE
US10604577B2 (en) 2015-10-22 2020-03-31 Allakos Inc. Methods and compositions for treating systemic mastocytosis
TW201723190A (en) 2015-10-22 2017-07-01 永斯醫療股份有限公司 Gene signatures for determining ICOS expression
MA44334A (en) 2015-10-29 2018-09-05 Novartis Ag ANTIBODY CONJUGATES INCLUDING A TOLL-TYPE RECEPTOR AGONIST
EP3184547A1 (en) 2015-10-29 2017-06-28 F. Hoffmann-La Roche AG Anti-tpbg antibodies and methods of use
WO2017072210A1 (en) 2015-10-29 2017-05-04 F. Hoffmann-La Roche Ag Anti-variant fc-region antibodies and methods of use
JO3555B1 (en) 2015-10-29 2020-07-05 Merck Sharp & Dohme Antibody neutralizing human respiratory syncytial virus
TW201730211A (en) 2015-10-30 2017-09-01 建南德克公司 Anti-Factor D antibodies and conjugates
KR102440160B1 (en) 2015-10-30 2022-09-02 제넨테크, 인크. Anti-HtrA1 antibodies and methods of use thereof
EP3368085B1 (en) 2015-11-01 2024-01-03 Massachusetts Institute Of Technology Modified alginates for anti-fibrotic materials and applications
WO2017079768A1 (en) 2015-11-08 2017-05-11 Genentech, Inc. Methods of screening for multispecific antibodies
KR20180082563A (en) 2015-11-19 2018-07-18 브리스톨-마이어스 스큅 컴퍼니 Antibodies to glucocorticoid-induced tumor necrosis factor receptor (GITR) and uses thereof
EP3383903A1 (en) 2015-11-30 2018-10-10 Bristol-Myers Squibb Company Anti human ip-10 antibodies and their uses
MX2018006477A (en) 2015-12-02 2018-09-03 Agenus Inc Antibodies and methods of use thereof.
EP3383908A1 (en) 2015-12-02 2018-10-10 Stsciences, Inc. Antibodies specific to glycosylated btla (b- and t- lymphocyte attenuator)
US10875920B2 (en) 2015-12-02 2020-12-29 Stcube & Co., Inc. Antibodies and molecules that immunospecifically bind to BTN1A1 and the therapeutic uses thereof
JP6794448B2 (en) 2015-12-04 2020-12-02 ノバルティス アーゲー Antibody Cytokine Graft Composition and Usage for Immunomodulation
US10954301B2 (en) 2015-12-14 2021-03-23 Macrogenics, Inc. Bispecific molecules having immunoreactivity with PD-1 and CTLA-4, and methods of use thereof
WO2017106129A1 (en) 2015-12-16 2017-06-22 Merck Sharp & Dohme Corp. Anti-lag3 antibodies and antigen-binding fragments
JP6797203B2 (en) 2015-12-17 2020-12-09 ザ・ジョンズ・ホプキンス・ユニバーシティー Improvement of systemic sclerosis by death receptor agonist
WO2017103895A1 (en) 2015-12-18 2017-06-22 Novartis Ag Antibodies targeting cd32b and methods of use thereof
KR102467124B1 (en) 2015-12-18 2022-11-15 추가이 세이야쿠 가부시키가이샤 Anti-c5 antibodies and methods of use
EP3862365A1 (en) 2016-01-08 2021-08-11 F. Hoffmann-La Roche AG Methods of treating cea-positive cancers using pd-1 axis binding antagonists and anti-cea/anti-cd3 bispecific antibodies
CN108602883A (en) 2016-01-20 2018-09-28 基因泰克公司 High-dose therapy for Alzheimer's disease
EP4035681A1 (en) 2016-01-28 2022-08-03 Institut National de la Santé et de la Recherche Médicale (INSERM) Methods and pharmaceutical composition for the treatment of cancer
ES2924741T3 (en) 2016-01-28 2022-10-10 Inst Nat Sante Rech Med Methods to Increase the Potency of Immune Checkpoint Inhibitors
WO2017129763A1 (en) 2016-01-28 2017-08-03 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for the treatment of signet ring cell gastric cancer
BR112018015659A2 (en) 2016-02-01 2018-12-26 Bioverativ Therapeutics Inc optimized factor viii genes
CN109071625A (en) 2016-02-04 2018-12-21 柯瑞斯公司 Smooth mutant and its application method
MX2018009800A (en) 2016-02-12 2018-11-09 Janssen Pharmaceutica Nv Anti-vista (b7h5) antibodies.
CN108699154A (en) 2016-02-26 2018-10-23 国家医疗保健研究所 There is the antibody and application thereof of specificity to BTLA
AU2017225854B2 (en) 2016-02-29 2020-11-19 Foundation Medicine, Inc. Therapeutic and diagnostic methods for cancer
AU2017228470A1 (en) 2016-03-04 2018-08-30 Bristol-Myers Squibb Company Combination therapy with anti-CD73 antibodies
JP6430025B2 (en) 2016-03-15 2018-11-28 中外製薬株式会社 Methods of treating cancer using PD-1 binding antagonists and anti-GPC3 antibodies
JP2019518713A (en) 2016-03-16 2019-07-04 メリマック ファーマシューティカルズ インコーポレーティッド Modified TRAIL for Cancer Therapy
KR102438140B1 (en) 2016-03-22 2022-08-31 엥스띠뛰 나씨오날 드 라 쌍떼 에 드 라 흐쉐르슈 메디깔 Humanized anti-claudin-1 antibodies and uses thereof
CN108697799A (en) 2016-03-22 2018-10-23 生态学有限公司 The application of anti-LGR5 monoclonal antibodies
EP4273551A3 (en) 2016-03-25 2024-01-17 F. Hoffmann-La Roche AG Multiplexed total antibody and antibody-conjugated drug quantification assay
MA45328A (en) 2016-04-01 2019-02-06 Avidity Biosciences Llc NUCLEIC ACID-POLYPEPTIDE COMPOSITIONS AND USES THEREOF
US11084879B2 (en) 2016-04-07 2021-08-10 The Johns Hopkins University Compositions and methods for treating pancreatitis and pain with death receptor agonists
EP3865511A1 (en) 2016-04-14 2021-08-18 F. Hoffmann-La Roche AG Anti-rspo3 antibodies and methods of use
EP3442562B1 (en) 2016-04-15 2022-09-21 Evive Biotechnology (Shanghai) Ltd An il-22 dimer for use in treating necrotizing enterocolitis
AU2017248766A1 (en) 2016-04-15 2018-11-01 Genentech, Inc. Methods for monitoring and treating cancer
SG11201808783XA (en) 2016-04-15 2018-11-29 Alpine Immune Sciences Inc Cd80 variant immunomodulatory proteins and uses thereof
EP3442579A4 (en) 2016-04-15 2019-12-18 Immunext Inc. Anti-human vista antibodies and use thereof
MX2018012492A (en) 2016-04-15 2019-06-06 Genentech Inc Methods for monitoring and treating cancer.
WO2017181148A2 (en) 2016-04-15 2017-10-19 Alpine Immune Sciences, Inc. Icos ligand variant immunomodulatory proteins and uses thereof
EP3443012A4 (en) 2016-04-15 2019-09-11 Bioatla, LLC Anti-axl antibodies, antibody fragments and their immunoconjugates and uses thereof
CA3020864A1 (en) 2016-04-15 2017-10-19 Macrogenics, Inc. Novel b7-h3-binding molecules, antibody drug conjugates thereof and methods of use thereof
US11514331B2 (en) 2016-04-27 2022-11-29 Massachusetts Institute Of Technology Sequence-controlled polymer random access memory storage
US11410746B2 (en) 2016-04-27 2022-08-09 Massachusetts Institute Of Technology Stable nanoscale nucleic acid assemblies and methods thereof
US11312766B2 (en) 2016-04-27 2022-04-26 Novartis Ag Antibodies against growth differentiation factor 15 and uses thereof
AU2017259869A1 (en) 2016-05-02 2018-09-27 F. Hoffmann-La Roche Ag The contorsbody - a single chain target binder
EA039084B1 (en) 2016-05-09 2021-12-01 Бристол-Майерс Сквибб Компани Tl1a antibodies and uses thereof
WO2017194554A1 (en) 2016-05-10 2017-11-16 Inserm (Institut National De La Sante Et De La Recherche Medicale) Combinations therapies for the treatment of cancer
JP7089483B2 (en) 2016-05-11 2022-06-22 エフ・ホフマン－ラ・ロシュ・アクチェンゲゼルシャフト Modified anti-tenascin antibody and usage
RS63698B1 (en) 2016-05-13 2022-11-30 Bioatla Inc Anti-ror2 antibodies, antibody fragments, their immunoconjugates and uses thereof
ES2858151T3 (en) 2016-05-20 2021-09-29 Hoffmann La Roche PROTAC-Antibody Conjugates and Procedures for Use
US20190292259A1 (en) 2016-05-24 2019-09-26 Inserm (Institut National De La Sante Et De La Recherche Medicale) Methods and pharmaceutical compositions for the treatment of non small cell lung cancer (nsclc) that coexists with chronic obstructive pulmonary disease (copd)
TW201802121A (en) 2016-05-25 2018-01-16 諾華公司 Reversal binding agents for anti-factor XI/XIa antibodies and uses thereof
EP3465221B1 (en) 2016-05-27 2020-07-22 H. Hoffnabb-La Roche Ag Bioanalytical method for the characterization of site-specific antibody-drug conjugates
KR102543118B1 (en) 2016-05-27 2023-06-14 아게누스 인코포레이티드 Anti-tim-3 antibodies and methods of use thereof
JP7043425B2 (en) 2016-06-06 2022-03-29 ジェネンテック， インコーポレイテッド Silvestrol Antibodies-Drug Conjugates and Usage
JP7231411B2 (en) 2016-06-15 2023-03-01 ノバルティス アーゲー Methods of treating diseases using inhibitors of bone morphogenetic protein 6 (BMP6)
EP3475298A1 (en) 2016-06-24 2019-05-01 H. Hoffnabb-La Roche Ag Anti-polyubiquitin multispecific antibodies
MA45554A (en) 2016-07-01 2019-05-08 Resolve Therapeutics Llc OPTIMIZED BINUCLEASE FUSIONS.
JP6983824B2 (en) 2016-07-04 2021-12-17 エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft New antibody format
EP3481393B1 (en) 2016-07-05 2021-04-14 Beigene, Ltd. Combination of a pd-1 antagonist and a raf inhibitor for treating cancer
CN109757103B (en) 2016-07-14 2024-01-02 百时美施贵宝公司 Antibodies against TIM3 and uses thereof
WO2018014260A1 (en) 2016-07-20 2018-01-25 Nanjing Legend Biotech Co., Ltd. Multispecific antigen binding proteins and methods of use thereof
WO2018022479A1 (en) 2016-07-25 2018-02-01 Biogen Ma Inc. Anti-hspa5 (grp78) antibodies and uses thereof
US11471488B2 (en) 2016-07-28 2022-10-18 Alpine Immune Sciences, Inc. CD155 variant immunomodulatory proteins and uses thereof
CN110088127A (en) 2016-07-28 2019-08-02 高山免疫科学股份有限公司 CD155 variant immune modulator and application thereof
US11834490B2 (en) 2016-07-28 2023-12-05 Alpine Immune Sciences, Inc. CD112 variant immunomodulatory proteins and uses thereof
MX2018015721A (en) 2016-07-29 2019-05-27 Chugai Pharmaceutical Co Ltd Bispecific antibody exhibiting increased alternative fviii-cofactor-function activity.
NL2017267B1 (en) 2016-07-29 2018-02-01 Aduro Biotech Holdings Europe B V Anti-pd-1 antibodies
US11186634B2 (en) 2016-07-29 2021-11-30 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies targeting tumor associated macrophages and uses thereof
JP7148493B2 (en) 2016-08-01 2022-10-05 ゾーマ （ユーエス） リミテッド ライアビリティ カンパニー Parathyroid hormone receptor 1 (PTH1R) antibodies and uses thereof
NL2017270B1 (en) 2016-08-02 2018-02-09 Aduro Biotech Holdings Europe B V New anti-hCTLA-4 antibodies
EP3494141A4 (en) 2016-08-03 2020-04-08 Bio-Techne Corporation Identification of vsig3/vista as a novel immune checkpoint and use thereof for immunotherapy
SG11201801024XA (en) 2016-08-05 2018-05-30 Chugai Pharmaceutical Co Ltd Therapeutic or preventive compositions for il-8-related diseases
WO2018027124A1 (en) 2016-08-05 2018-02-08 Medimmune, Llc Anti-o2 antibodies and uses thereof
JP2019530434A (en) 2016-08-05 2019-10-24 ジェネンテック， インコーポレイテッド Multivalent and multi-epitope antibodies with agonist activity and methods of use
WO2018027203A1 (en) 2016-08-05 2018-02-08 Allakos, Inc. Anti-siglec-7 antibodies for the treatment of cancer
WO2018029124A1 (en) 2016-08-08 2018-02-15 F. Hoffmann-La Roche Ag Therapeutic and diagnostic methods for cancer
US10696722B2 (en) 2016-08-10 2020-06-30 Ajou University Industry-Academic Cooperation Foundation Heterodimeric Fc-fused cytokine and pharmaceutical composition comprising the same
CN109689111B (en) 2016-08-11 2024-04-05 基因泰克公司 Pyrrolobenzodiazepine prodrugs and antibody conjugates thereof
FI3500299T3 (en) 2016-08-19 2024-02-14 Beigene Switzerland Gmbh Combination of zanubrutinib with an anti-cd20 or an anti-pd-1 antibody for use in treating cancer
WO2018044970A1 (en) 2016-08-31 2018-03-08 University Of Rochester Human monoclonal antibodies to human endogenous retrovirus k envelope (herv-k) and uses thereof
WO2018041989A1 (en) 2016-09-02 2018-03-08 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for diagnosing and treating refractory celiac disease type 2
US20190270821A1 (en) 2016-09-13 2019-09-05 Humanigen, Inc. Epha3 antibodies for the treatment of pulmonary fibrosis
US11008402B2 (en) 2016-09-15 2021-05-18 Universitat Stuttgart Antigen binding protein against HER3
BR112019005129A2 (en) 2016-09-16 2019-06-04 Shanghai Henlius Biotech Inc anti-pd-1 antibodies
SG10201607778XA (en) 2016-09-16 2018-04-27 Chugai Pharmaceutical Co Ltd Anti-Dengue Virus Antibodies, Polypeptides Containing Variant Fc Regions, And Methods Of Use
SG11201901950TA (en) 2016-09-19 2019-04-29 Celgene Corp Methods of treating immune disorders using pd-1 binding proteins
WO2018050878A1 (en) 2016-09-19 2018-03-22 F. Hoffmann-La Roche Ag Complement factor based affinity chromatography
JP2019534859A (en) 2016-09-19 2019-12-05 セルジーン コーポレイション Method for treating vitiligo using PD-1 binding protein
KR20240035625A (en) 2016-09-21 2024-03-15 넥스트큐어 인코포레이티드 Antibodies for siglec-15 and methods of use thereof
UA124269C2 (en) 2016-09-23 2021-08-18 Дженентек, Інк. Uses of il-13 antagonists for treating atopic dermatitis
JOP20190055A1 (en) 2016-09-26 2019-03-24 Merck Sharp & Dohme Anti-cd27 antibodies
MX2019003543A (en) 2016-09-28 2019-06-17 Xoma Us Llc Antibodies that bind interleukin-2 and uses thereof.
EP3522933B1 (en) 2016-10-05 2021-12-15 F. Hoffmann-La Roche AG Methods for preparing antibody drug conjugates
KR20190072528A (en) 2016-10-06 2019-06-25 제넨테크, 인크. Treatment and Diagnosis Methods for Cancer
US10844119B2 (en) 2016-10-11 2020-11-24 Agenus Inc. Anti-LAG-3 antibodies and methods of use thereof
WO2018068201A1 (en) 2016-10-11 2018-04-19 Nanjing Legend Biotech Co., Ltd. Single-domain antibodies and variants thereof against ctla-4
KR20190064636A (en) 2016-10-19 2019-06-10 메디뮨 엘엘씨 Anti-O1 antibodies and uses thereof
US11186635B2 (en) 2016-10-25 2021-11-30 Institut National De La Santé Et De La Recherché Médical (Inserm) Monoclonal antibodies binding to the CD160 transmembrane isoform
EP3532091A2 (en) 2016-10-29 2019-09-04 H. Hoffnabb-La Roche Ag Anti-mic antibidies and methods of use
TWI788307B (en) 2016-10-31 2023-01-01 美商艾歐凡斯生物治療公司 Engineered artificial antigen presenting cells for tumor infiltrating lymphocyte expansion
MX2019004621A (en) 2016-11-02 2019-11-28 Engmab Sarl Bispecific antibody against bcma and cd3 and an immunological drug for combined use in treating multiple myeloma.
MD3535298T2 (en) 2016-11-02 2022-01-31 Jounce Therapeutics Inc Antibodies to pd-1 and uses thereof
US11339209B2 (en) 2016-11-14 2022-05-24 Novartis Ag Compositions, methods, and therapeutic uses related to fusogenic protein minion
CA3042435A1 (en) 2016-11-15 2018-05-24 Genentech, Inc. Dosing for treatment with anti-cd20/anti-cd3 bispecific antibodies
JOP20190100A1 (en) 2016-11-19 2019-05-01 Potenza Therapeutics Inc Anti-gitr antigen-binding proteins and methods of use thereof
AU2017361887B2 (en) 2016-11-21 2019-08-15 Cureab Gmbh Anti-GP73 antibodies and immunoconjugates
MA46893A (en) 2016-11-23 2019-10-02 Bioverativ Therapeutics Inc BISPECIFIC ANTIBODIES BINDING TO COAGULATION FACTOR IX AND COAGULATION FACTOR X
MX2019006446A (en) 2016-12-02 2019-12-11 Bioverativ Therapeutics Inc Methods of inducing immune tolerance to clotting factors.
MA46967A (en) 2016-12-02 2019-10-09 Bioverativ Therapeutics Inc METHODS OF TREATMENT OF HEMOPHILIC ARTHROPATHY USING CHEMERICAL COAGULATION FACTORS
CN110300599A (en) 2016-12-07 2019-10-01 艾吉纳斯公司 Antibody and its application method
WO2018106781A1 (en) 2016-12-07 2018-06-14 Genentech, Inc Anti-tau antibodies and methods of use
WO2018106776A2 (en) 2016-12-07 2018-06-14 Genentech, Inc. Anti-tau antibodies and methods of use
IL307242A (en) 2016-12-07 2023-11-01 Agenus Inc Anti-ctla-4 antibodies and methods of use thereof
KR20190095921A (en) 2016-12-12 2019-08-16 제넨테크, 인크. How to Treat Cancer Using Anti-PD-L1 Antibody and Antiandrogen
CA3039446A1 (en) 2016-12-20 2018-06-28 F. Hoffmann-La Roche Ag Combination therapy of anti-cd20/anti-cd3 bispecific antibodies and 4-1bb (cd137) agonists
WO2018116255A1 (en) 2016-12-23 2018-06-28 Novartis Ag Factor xi antibodies and methods of use
JOP20190134A1 (en) 2016-12-23 2019-06-02 Potenza Therapeutics Inc Anti-neuropilin antigen-binding proteins and methods of use thereof
US20180230218A1 (en) 2017-01-04 2018-08-16 Immunogen, Inc. Met antibodies and immunoconjugates and uses thereof
MX2019008199A (en) 2017-01-06 2019-11-25 Avidity Biosciences Llc Nucleic acid-polypeptide compositions and methods of inducing exon skipping.
MX2019007963A (en) 2017-01-06 2019-10-21 Iovance Biotherapeutics Inc Expansion of tumor infiltrating lymphocytes (tils) with tumor necrosis factor receptor superfamily (tnfrsf) agonists and therapeutic combinations of tils and tnfrsf agonists.
JP2020503351A (en) 2017-01-06 2020-01-30 アイオバンス バイオセラピューティクス，インコーポレイテッド Proliferation of tumor infiltrating lymphocytes by potassium channel agonist and its therapeutic use
US20180244785A1 (en) 2017-01-09 2018-08-30 Merrimack Pharmaceuticals, Inc. Anti-fgfr antibodies and methods of use
WO2018132597A1 (en) 2017-01-12 2018-07-19 Eureka Therapeutics, Inc. Constructs targeting histone h3 peptide/mhc complexes and uses thereof
US11555038B2 (en) 2017-01-25 2023-01-17 Beigene, Ltd. Crystalline forms of (S)-7-(1-(but-2-ynoyl)piperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide, preparation, and uses thereof
SG10201908697XA (en) 2017-01-31 2019-10-30 Chugai Pharmaceutical Co Ltd A pharmaceutical composition for use in the treatment or prevention of a c5-related disease and a method for treating or preventing a c5-related disease
JOP20190187A1 (en) 2017-02-03 2019-08-01 Novartis Ag Anti-ccr7 antibody drug conjugates
AU2018218557B9 (en) 2017-02-08 2021-06-24 Novartis Ag FGF21 mimetic antibodies and uses thereof
JP6995127B2 (en) 2017-02-10 2022-02-04 ジェネンテック， インコーポレイテッド Anti-tryptase antibody, its composition, and its use
CA3051839A1 (en) 2017-02-17 2018-08-23 Bristol-Myers Squibb Company Antibodies to alpha-synuclein and uses thereof
WO2018156180A1 (en) 2017-02-24 2018-08-30 Kindred Biosciences, Inc. Anti-il31 antibodies for veterinary use
CA3052670A1 (en) 2017-03-01 2018-09-07 Genentech, Inc. Diagnostic and therapeutic methods for cancer
JP2020510432A (en) 2017-03-02 2020-04-09 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル Antibodies with specificity for NECTIN-4 and uses thereof
EP3595645A1 (en) 2017-03-15 2020-01-22 INSERM (Institut National de la Santé et de la Recherche Médicale) Pharmaceutical compositions for the treatment of thrombosis in patients suffering from a myeloproliferative neoplasm
ES2963638T3 (en) 2017-03-16 2024-04-01 Alpine Immune Sciences Inc Immunomodulatory proteins of PD-L1 variants and their uses
WO2018170023A1 (en) 2017-03-16 2018-09-20 Alpine Immune Sciences, Inc. Pd-l2 variant immunomodulatory proteins and uses thereof
NZ756395A (en) 2017-03-16 2024-01-26 Alpine Immune Sciences Inc Cd80 variant immunomodulatory proteins and uses thereof
JP7227151B2 (en) 2017-03-22 2023-02-21 ジェネンテック， インコーポレイテッド Antibody Compositions Optimized for Treatment of Eye Disorders
CA3054837A1 (en) 2017-03-24 2018-09-27 Novartis Ag Methods for preventing and treating heart disease
AU2018244224A1 (en) 2017-03-28 2019-09-19 Genentech, Inc. Methods of treating neurodegenerative diseases
JOP20190203A1 (en) 2017-03-30 2019-09-03 Potenza Therapeutics Inc Anti-tigit antigen-binding proteins and methods of use thereof
WO2018187191A1 (en) 2017-04-03 2018-10-11 Jounce Therapeutics, Inc Compositions and methods for the treatment of cancer
WO2018185618A1 (en) 2017-04-03 2018-10-11 Novartis Ag Anti-cdh6 antibody drug conjugates and anti-gitr antibody combinations and methods of treatment
PL3606954T3 (en) 2017-04-05 2022-10-17 F. Hoffmann-La Roche Ag Anti-lag3 antibodies
TWI796329B (en) 2017-04-07 2023-03-21 美商默沙東有限責任公司 Anti-ilt4 antibodies and antigen-binding fragments
EP3609922A2 (en) 2017-04-13 2020-02-19 Aduro Biotech Holdings, Europe B.V. Anti-sirp alpha antibodies
JP7297672B2 (en) 2017-04-13 2023-06-26 アジェナス インコーポレイテッド ANTI-CD137 ANTIBODY AND METHODS OF USE THEREOF
CA3060080C (en) 2017-04-14 2024-01-23 The United States Of America As Represented By The Department Of Veterans Affairs Compositions and methods for treating pulmonary fibrosis
JP7289420B6 (en) 2017-04-14 2023-06-30 エクセリクシス， インク． AMHRII-binding compounds for preventing or treating lung cancer
CN110621787A (en) 2017-04-14 2019-12-27 豪夫迈·罗氏有限公司 Methods for diagnosis and treatment of cancer
CA3058282A1 (en) 2017-04-14 2018-10-18 Gamamabs Pharma Amhrii-binding compounds for preventing or treating cancers
PE20200150A1 (en) 2017-04-21 2020-01-17 Genentech Inc USE OF KLK5 ANTAGONISTS FOR THE TREATMENT OF A DISEASE
SG11201909728XA (en) 2017-04-26 2019-11-28 Eureka Therapeutics Inc Constructs specifically recognizing glypican 3 and uses thereof
JP2020517695A (en) 2017-04-27 2020-06-18 テサロ， インコーポレイテッド Antibody drugs against lymphocyte activation gene-3 (LAG-3) and their use
AR111651A1 (en) 2017-04-28 2019-08-07 Novartis Ag CONJUGATES OF ANTIBODIES THAT INCLUDE TOLL TYPE RECEIVER AGONISTS AND COMBINATION THERAPIES
HUE062927T2 (en) 2017-05-01 2023-12-28 Agenus Inc Anti-tigit antibodies and methods of use thereof
WO2018204594A1 (en) 2017-05-04 2018-11-08 Acceleron Pharma Inc. Tgf-beta receptor type ii fusion proteins and uses thereof
MX2019013137A (en) 2017-05-05 2020-07-14 Allakos Inc Methods and compositions for treating allergic ocular diseases.
WO2018209115A1 (en) 2017-05-10 2018-11-15 Iovance Biotherapeutics, Inc. Expansion of tumor infiltrating lymphocytes from liquid tumors and therapeutic uses thereof
WO2019103857A1 (en) 2017-11-22 2019-05-31 Iovance Biotherapeutics, Inc. Expansion of peripheral blood lymphocytes (pbls) from peripheral blood
WO2018209125A1 (en) 2017-05-10 2018-11-15 Fred Hutchinson Cancer Research Center Epstein barr virus antibodies, vaccines, and uses of the same
US11168129B2 (en) 2017-05-15 2021-11-09 University Of Rochester Broadly neutralizing anti-influenza human monoclonal antibody and uses thereof
WO2018215937A1 (en) 2017-05-24 2018-11-29 Novartis Ag Interleukin-7 antibody cytokine engrafted proteins and methods of use in the treatment of cancer
CN110662762A (en) 2017-05-24 2020-01-07 诺华股份有限公司 Antibody cytokine transplantation proteins and methods for treating cancer
EP3630162A1 (en) 2017-05-24 2020-04-08 Novartis AG Antibody-cytokine engrafted proteins and methods of use
JOP20190271A1 (en) 2017-05-24 2019-11-21 Novartis Ag Antibody-cytokine engrafted proteins and methods of use for immune related disorders
CN116333129A (en) 2017-05-25 2023-06-27 百时美施贵宝公司 Antibodies comprising modified heavy chain constant regions
EP3630835A1 (en) 2017-05-31 2020-04-08 STCube & Co., Inc. Antibodies and molecules that immunospecifically bind to btn1a1 and the therapeutic uses thereof
KR20200024158A (en) 2017-05-31 2020-03-06 주식회사 에스티큐브앤컴퍼니 How to treat cancer using antibodies and molecules that immunospecifically bind to BTN1A1
AU2018276140A1 (en) 2017-06-01 2019-12-19 Compugen Ltd. Triple combination antibody therapies
KR20200026209A (en) 2017-06-06 2020-03-10 주식회사 에스티큐브앤컴퍼니 How to treat cancer using antibodies and molecules that bind BTN1A1 or BTN1A1-ligand
UY37758A (en) 2017-06-12 2019-01-31 Novartis Ag METHOD OF MANUFACTURING OF BIESPECTIFIC ANTIBODIES, BISPECTIFIC ANTIBODIES AND THERAPEUTIC USE OF SUCH ANTIBODIES
WO2018229715A1 (en) 2017-06-16 2018-12-20 Novartis Ag Compositions comprising anti-cd32b antibodies and methods of use thereof
WO2018229706A1 (en) 2017-06-16 2018-12-20 Novartis Ag Combination therapy for the treatment of cancer
WO2018236904A1 (en) 2017-06-19 2018-12-27 Surface Oncology, Inc. Combination of anti-cd47 antibodies and cell death-inducing agents, and uses thereof
CA3070991C (en) 2017-06-19 2023-10-17 Massachusetts Institute Of Technology Automated methods for scalable, parallelized enzymatic biopolymer synthesis and modification using microfluidic devices
EP3642239A1 (en) 2017-06-20 2020-04-29 Amgen Inc. Method of treating or ameliorating metabolic disorders using binding proteins for gastric inhibitory peptide receptor (gipr) in combination with glp-1 agonists
WO2018234843A1 (en) 2017-06-22 2018-12-27 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for the treatment of fibrosis with agents capable of inhibiting the activation of mucosal-associated invariant t (mait) cells
US11597768B2 (en) 2017-06-26 2023-03-07 Beigene, Ltd. Immunotherapy for hepatocellular carcinoma
CA3063659A1 (en) 2017-06-28 2019-01-03 Novartis Ag Methods for preventing and treating urinary incontinence
EP3652206A1 (en) 2017-07-10 2020-05-20 International-Drug-Development-Biotech Treatment of b cell malignancies using afucosylated pro-apoptotic anti-cd19 antibodies in combination with anti cd20 antibodies or chemotherapeutics
JP7402521B2 (en) 2017-07-11 2023-12-21 コンパス セラピューティクス リミテッド ライアビリティ カンパニー Agonistic antibodies that bind to human CD137 and uses thereof
TWI823859B (en) 2017-07-21 2023-12-01 美商建南德克公司 Therapeutic and diagnostic methods for cancer
GB201711809D0 (en) 2017-07-21 2017-09-06 Governors Of The Univ Of Alberta Antisense oligonucleotide
EP3658173A1 (en) 2017-07-25 2020-06-03 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for modulating monocytopoiesis
AU2018308364C1 (en) 2017-07-26 2023-02-16 Forty Seven, Inc. Anti-SIRP-alpha antibodies and related methods
SI3658184T1 (en) 2017-07-27 2024-01-31 Alexion Pharmaceuticals, Inc., High concentration anti-c5 antibody formulations
JP7374883B2 (en) 2017-08-09 2023-11-07 バイオベラティブ セラピューティクス インコーポレイテッド Nucleic acid molecules and their uses
EP3668536A4 (en) * 2017-08-15 2021-05-26 Kindred Biosciences, Inc. Igg fc variants for veterinary use
WO2019036605A2 (en) 2017-08-17 2019-02-21 Massachusetts Institute Of Technology Multiple specificity binders of cxc chemokines and uses thereof
MX2020002070A (en) 2017-08-22 2020-03-24 Sanabio Llc Soluble interferon receptors and uses thereof.
MA49950A (en) 2017-08-25 2020-07-01 Five Prime Therapeutics Inc ANTI-B7-H4 ANTIBODIES AND PROCEDURES FOR USE
CA3074111A1 (en) 2017-08-28 2019-03-07 Angiex, Inc. Anti-tm4sf1 antibodies and methods of using same
MX2020002612A (en) 2017-09-07 2020-07-13 Univ Res Inst Inc Augusta Antibodies to programmed cell death protein 1.
WO2019059411A1 (en) 2017-09-20 2019-03-28 Chugai Seiyaku Kabushiki Kaisha Dosage regimen for combination therapy using pd-1 axis binding antagonists and gpc3 targeting agent
WO2019057742A1 (en) 2017-09-20 2019-03-28 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for modulating autophagy
BR112020005671A2 (en) 2017-09-21 2020-10-20 INSERM (Institut National de la Santé et de la Recherche Médicale) antibodies that have specificity for btn2 and their uses
WO2019075090A1 (en) 2017-10-10 2019-04-18 Tilos Therapeutics, Inc. Anti-lap antibodies and uses thereof
CA3198255A1 (en) 2017-10-10 2019-04-18 Alpine Immune Sciences, Inc. Ctla-4 variant immunomodulatory proteins and uses thereof
WO2019075270A1 (en) 2017-10-12 2019-04-18 Amesino Llc Vegfr-antibody light chain fusion protein
BR112020007309A2 (en) 2017-10-14 2020-09-29 Cytomx Therapeutics, Inc. antibodies, activable antibodies, bispecific antibodies and bispecific activable antibodies and methods of using them
WO2019079520A2 (en) 2017-10-18 2019-04-25 Alpine Immune Sciences, Inc. Variant icos ligand immunomodulatory proteins and related compositions and methods
TW201932142A (en) 2017-10-20 2019-08-16 瑞士商赫孚孟拉羅股份公司 Method for generating multispecific antibodies from monospecific antibodies
US20210040205A1 (en) 2017-10-25 2021-02-11 Novartis Ag Antibodies targeting cd32b and methods of use thereof
MX2020004100A (en) 2017-10-30 2020-07-24 Hoffmann La Roche Method for in vivo generation of multispecific antibodies from monospecific antibodies.
WO2019089753A2 (en) 2017-10-31 2019-05-09 Compass Therapeutics Llc Cd137 antibodies and pd-1 antagonists and uses thereof
AU2018359967A1 (en) 2017-11-06 2020-04-23 Genentech, Inc. Diagnostic and therapeutic methods for cancer
WO2019094928A1 (en) 2017-11-10 2019-05-16 Massachusetts Institute Of Technology Microbial production of pure single stranded nucleic acids
EP3713961A2 (en) 2017-11-20 2020-09-30 Compass Therapeutics LLC Cd137 antibodies and tumor antigen-targeting antibodies and uses thereof
EP3713965A1 (en) 2017-11-22 2020-09-30 Novartis AG Reversal binding agents for anti-factor xi/xia antibodies and uses thereof
WO2019101995A1 (en) 2017-11-27 2019-05-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for cardiac regeneration
US11786529B2 (en) 2017-11-29 2023-10-17 Beigene Switzerland Gmbh Treatment of indolent or aggressive B-cell lymphomas using a combination comprising BTK inhibitors
JP2021508439A (en) 2017-12-01 2021-03-11 ノバルティス アーゲー Polyomavirus neutralizing antibody
SG11202003930YA (en) 2017-12-01 2020-05-28 Pfizer Anti-cxcr5 antibodies and compositions and uses thereof
EP3720448A4 (en) 2017-12-06 2021-11-03 Avidity Biosciences, Inc. Compositions and methods of treating muscle atrophy and myotonic dystrophy
EP3720877A1 (en) 2017-12-08 2020-10-14 Argenx BVBA Use of fcrn antagonists for treatment of generalized myasthenia gravis
WO2019118873A2 (en) 2017-12-15 2019-06-20 Iovance Biotherapeutics, Inc. Systems and methods for determining the beneficial administration of tumor infiltrating lymphocytes, and methods of use thereof and beneficial administration of tumor infiltrating lymphocytes, and methods of use thereof
AU2018393073A1 (en) 2017-12-19 2020-07-02 Surrozen Operating, Inc. Wnt surrogate molecules and uses thereof
JP7317016B2 (en) 2017-12-19 2023-07-28 スロゼン オペレーティング， インコーポレイテッド Anti-LRP5/6 Antibodies and Methods of Use
WO2019126536A1 (en) 2017-12-20 2019-06-27 Alexion Pharmaceuticals Inc. Humanized anti-cd200 antibodies and uses thereof
WO2019126133A1 (en) 2017-12-20 2019-06-27 Alexion Pharmaceuticals, Inc. Liquid formulations of anti-cd200 antibodies
WO2019126472A1 (en) 2017-12-22 2019-06-27 Genentech, Inc. Use of pilra binding agents for treatment of a disease
JP7391868B2 (en) 2017-12-22 2023-12-05 ジョウンセ セラピューティクス， インク． Antibody against LILRB2
KR20200104886A (en) 2017-12-28 2020-09-04 난징 레전드 바이오테크 씨오., 엘티디. Antibodies and variants against PD-L1
US11905327B2 (en) 2017-12-28 2024-02-20 Nanjing Legend Biotech Co., Ltd. Single-domain antibodies and variants thereof against TIGIT
EP3731864A1 (en) 2017-12-29 2020-11-04 F. Hoffmann-La Roche SA Anti-vegf antibodies and methods of use
CA3087149A1 (en) 2018-01-03 2019-07-11 Alpine Immune Sciences, Inc. Multi-domain immunomodulatory proteins and methods of use thereof
KR20200118029A (en) 2018-01-04 2020-10-14 아이코닉 테라퓨틱스, 인코포레이티드 Anti-tissue factor antibodies, antibody-drug conjugates, and related methods
KR20200106525A (en) 2018-01-05 2020-09-14 에이씨 이뮨 에스.에이. Misfolded TDP-43 binding molecule
US20220089720A1 (en) 2018-01-12 2022-03-24 Bristol-Myers Squibb Company Antibodies against tim3 and uses thereof
SG11202004233UA (en) 2018-01-15 2020-06-29 Nanjing Legend Biotech Co Ltd Single-domain antibodies and variants thereof against pd-1
US20200339686A1 (en) 2018-01-16 2020-10-29 Lakepharma, Inc. Bispecific antibody that binds cd3 and another target
US11078283B2 (en) 2018-01-17 2021-08-03 Apexigen, Inc. Anti-PD-L1 antibodies and methods of use
CN112566934A (en) 2018-01-23 2021-03-26 奈斯科尔公司 B7-H4 antibodies and methods of use thereof
WO2019145413A1 (en) 2018-01-25 2019-08-01 INSERM (Institut National de la Santé et de la Recherche Médicale) Antagonists of il-33 for use in methods for preventing ischemia reperfusion injury in an organ
RU2020127792A (en) 2018-01-26 2022-02-28 Дженентек, Инк. IL-22-Fc fusion proteins AND METHODS OF APPLICATION
LT3743088T (en) 2018-01-26 2022-12-27 F. Hoffmann-La Roche Ag Il-22 fc compositions and methods of use
JP7438953B2 (en) 2018-02-01 2024-02-27 イノベント バイオロジックス （スウツォウ） カンパニー，リミテッド Fully humanized anti-B cell maturation antigen (BCMA) single chain antibody and its applications
CN111918674A (en) 2018-02-01 2020-11-10 比奥维拉迪维治疗股份有限公司 Use of lentiviral vectors expressing factor VIII
WO2019148412A1 (en) 2018-02-01 2019-08-08 Merck Sharp & Dohme Corp. Anti-pd-1/lag3 bispecific antibodies
CN111868082A (en) 2018-02-02 2020-10-30 博奥泰克尼公司 Compounds that modulate the interaction of VISTA and VSIG3 and methods of making and using the same
MA51793A (en) 2018-02-08 2020-12-16 Hoffmann La Roche BISPECIFIC ANTIGEN BINDING MOLECULES AND METHODS OF USE
JP7418337B2 (en) 2018-02-09 2024-01-19 ジェネンテック， インコーポレイテッド Treatment and diagnosis of mast cell-mediated inflammatory diseases
JP2021512962A (en) 2018-02-13 2021-05-20 アイオバンス バイオセラピューティクス，インコーポレイテッド Expansion culture of tumor-infiltrating lymphocytes (TIL) with adenosine A2A receptor antagonist and therapeutic combination of TIL and adenosine A2A receptor antagonist
JP7350756B2 (en) 2018-02-14 2023-09-26 アバ セラピューティクス アーゲー Anti-human PD-L2 antibody
SG11202007694UA (en) 2018-02-21 2020-09-29 Genentech Inc DOSING FOR TREATMENT WITH IL-22 Fc FUSION PROTEINS
TW202000702A (en) 2018-02-26 2020-01-01 美商建南德克公司 Dosing for treatment with anti-TIGIT and anti-PD-L1 antagonist antibodies
WO2019169229A1 (en) 2018-03-01 2019-09-06 Nextcure, Inc. Klrg1 binding compositions and methods of use thereof
NL2020520B1 (en) 2018-03-02 2019-09-12 Labo Bio Medical Invest B V Multispecific binding molecules for the prevention, treatment and diagnosis of neurodegenerative disorders
WO2019169212A1 (en) 2018-03-02 2019-09-06 Five Prime Therapeutics, Inc. B7-h4 antibodies and methods of use thereof
EP3765489A1 (en) 2018-03-13 2021-01-20 F. Hoffmann-La Roche AG Therapeutic combination of 4-1 bb agonists with anti-cd20 antibodies
WO2019178269A2 (en) 2018-03-14 2019-09-19 Surface Oncology, Inc. Antibodies that bind cd39 and uses thereof
US20200040103A1 (en) 2018-03-14 2020-02-06 Genentech, Inc. Anti-klk5 antibodies and methods of use
CN112119090B (en) 2018-03-15 2023-01-13 中外制药株式会社 Anti-dengue virus antibodies cross-reactive to Zika virus and methods of use
SG11202008593PA (en) 2018-03-21 2020-10-29 Five Prime Therapeutics Inc ANTIBODIES BINDING TO VISTA AT ACIDIC pH
KR20210005007A (en) 2018-03-22 2021-01-13 서피스 온콜로지, 인크. Anti-IL-27 antibodies and uses thereof
AU2019236865A1 (en) 2018-03-23 2020-10-01 Bristol-Myers Squibb Company Antibodies against MICA and/or MICB and uses thereof
EA202092316A1 (en) 2018-03-28 2021-05-25 Бристол-Маерс Сквибб Компани FUSION PROTEINS OF INTERLEUKIN-2 / ALPHA-RECEPTOR OF INTERLEUKIN-2 AND METHODS OF APPLICATION
WO2019190984A1 (en) 2018-03-29 2019-10-03 Pfizer Inc. Lfa3 variants and compositions and uses thereof
CN111936625A (en) 2018-03-29 2020-11-13 豪夫迈·罗氏有限公司 Modulating lactogenic activity in mammalian cells
SG11202006674QA (en) 2018-03-30 2020-08-28 Nanjing Legend Biotech Co Ltd Single-domain antibodies against lag-3 and uses thereof
SG11202009625WA (en) 2018-04-02 2020-10-29 Bristol Myers Squibb Co Anti-trem-1 antibodies and uses thereof
WO2019193375A1 (en) 2018-04-04 2019-10-10 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of fzd7 inhibitors for the treatment of retinal neovascularization
TW202011029A (en) 2018-04-04 2020-03-16 美商建南德克公司 Methods for detecting and quantifying FGF21
EP3552631A1 (en) 2018-04-10 2019-10-16 Inatherys Antibody-drug conjugates and their uses for the treatment of cancer
WO2019200357A1 (en) 2018-04-12 2019-10-17 Surface Oncology, Inc. Biomarker for cd47 targeting therapeutics and uses therefor
US11648317B2 (en) 2018-04-13 2023-05-16 Genentech, Inc. Stable anti-CD79B immunoconjugate formulations
US20210070871A1 (en) 2018-04-25 2021-03-11 Prometheus Biosciences, Inc. Optimized anti-tl1a antibodies
WO2019207030A1 (en) 2018-04-26 2019-10-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for predicting a response with an immune checkpoint inhibitor in a patient suffering from a lung cancer
US11958895B2 (en) 2018-05-03 2024-04-16 University Of Rochester Anti-influenza neuraminidase monoclonal antibodies and uses thereof
CN110464842B (en) 2018-05-11 2022-10-14 信达生物制药(苏州)有限公司 Formulations comprising anti-PCSK 9 antibodies and uses thereof
BR112020022164A2 (en) 2018-05-18 2021-02-02 Bioverativ Therapeutics Inc. methods of treating hemophilia a
WO2019226617A1 (en) 2018-05-21 2019-11-28 Compass Therapeutics Llc Compositions and methods for enhancing the killing of target cells by nk cells
WO2019226658A1 (en) 2018-05-21 2019-11-28 Compass Therapeutics Llc Multispecific antigen-binding compositions and methods of use
TW202015726A (en) 2018-05-30 2020-05-01 瑞士商諾華公司 Entpd2 antibodies, combination therapies, and methods of using the antibodies and combination therapies
CN112165974A (en) 2018-05-31 2021-01-01 诺华股份有限公司 Hepatitis B antibodies
EP3802611A2 (en) 2018-06-01 2021-04-14 Novartis AG Binding molecules against bcma and uses thereof
KR20210027352A (en) 2018-06-04 2021-03-10 바이오젠 엠에이 인코포레이티드 Anti-VLA-4 antibody with reduced effector function
WO2019235426A1 (en) 2018-06-04 2019-12-12 中外製薬株式会社 Antigen-binding molecule showing changed half-life in cytoplasm
AU2019282264A1 (en) 2018-06-05 2020-11-26 Amgen Inc. Modulating antibody dependent cellular phagocytosis
US20210363219A1 (en) 2018-06-15 2021-11-25 Alpine Immune Sciences, Inc. Pd-1 variant immunomodulatory proteins and uses thereof
US20200030443A1 (en) 2018-06-23 2020-01-30 Genentech, Inc. Methods of treating lung cancer with a pd-1 axis binding antagonist, a platinum agent, and a topoisomerase ii inhibitor
BR112020026512A2 (en) 2018-07-03 2021-04-06 Bristol-Myers Squibb Company FGF-21 FORMULATIONS
BR112020027095A2 (en) 2018-07-09 2021-03-30 Five Prime Therapeutics, Inc. ILT4 BINDING ANTIBODIES
WO2020014306A1 (en) 2018-07-10 2020-01-16 Immunogen, Inc. Met antibodies and immunoconjugates and uses thereof
KR20210031722A (en) 2018-07-11 2021-03-22 파이브 프라임 테라퓨틱스, 인크. Antibodies that bind to VISTA at acidic pH
KR20210034622A (en) 2018-07-18 2021-03-30 제넨테크, 인크. Lung cancer treatment method using PD-1 axis binding antagonist, anti-metabolite, and platinum agent
JP7072715B2 (en) 2018-07-20 2022-05-20 サーフィス オンコロジー インコーポレイテッド Anti-CD112R Compositions and Methods
TWI754157B (en) 2018-07-25 2022-02-01 大陸商信達生物製藥（蘇州）有限公司 Anti-tigit antibody and use thereof
WO2020025703A1 (en) 2018-08-01 2020-02-06 Imcheck Therapeutics Sas Anti-btn3a antibodies and their use in treating cancer or infectious disorders
CN112512563A (en) 2018-08-01 2021-03-16 中外制药株式会社 Pharmaceutical composition for treating or preventing C5-related diseases and method for treating or preventing C5-related diseases
KR20210040989A (en) 2018-08-03 2021-04-14 추가이 세이야쿠 가부시키가이샤 Antigen-binding molecule comprising two antigen-binding domains linked to each other
MA50586A (en) 2018-08-09 2020-09-16 Regeneron Pharma METHODS FOR EVALUATING THE BINDING AFFINITY OF AN ANTIBODY VARIANT TO THE NEONATAL FC RECEPTOR
WO2020033926A2 (en) 2018-08-09 2020-02-13 Compass Therapeutics Llc Antibodies that bind cd277 and uses thereof
EP3833443A1 (en) 2018-08-09 2021-06-16 Compass Therapeutics LLC Antigen binding agents that bind cd277 and uses thereof
JP2021533753A (en) 2018-08-09 2021-12-09 バイオベラティブ セラピューティクス インコーポレイテッド Nucleic acid molecules and their use for non-viral gene therapy
WO2020033925A2 (en) 2018-08-09 2020-02-13 Compass Therapeutics Llc Antibodies that bind cd277 and uses thereof
EA202190451A1 (en) 2018-08-10 2021-07-13 Чугаи Сейяку Кабусики Кайся ANTI-CD137 ANTIGEN-BINDING MOLECULES AND THEIR APPLICATION
AU2019319970A1 (en) 2018-08-10 2021-03-11 Genentech, Inc. Cell culture strategies for modulating protein glycosylation
TW202021618A (en) 2018-08-17 2020-06-16 美商２３與我有限公司 Anti-il1rap antibodies and methods of use thereof
TW202031273A (en) 2018-08-31 2020-09-01 美商艾歐凡斯生物治療公司 Treatment of nsclc patients refractory for anti-pd-1 antibody
GB201814281D0 (en) 2018-09-03 2018-10-17 Femtogenix Ltd Cytotoxic agents
EP3847650A1 (en) 2018-09-06 2021-07-14 The Broad Institute, Inc. Nucleic acid assemblies for use in targeted delivery
SG11202100888WA (en) 2018-09-07 2021-02-25 Pfizer Anti-avb8 antibodies and compositions and uses thereof
WO2020053742A2 (en) 2018-09-10 2020-03-19 Novartis Ag Anti-hla-hbv peptide antibodies
JP2022500371A (en) 2018-09-11 2022-01-04 アムジエン・インコーポレーテツド How to regulate antibody-dependent cellular cytotoxicity
WO2020056170A1 (en) 2018-09-12 2020-03-19 Fred Hutchinson Cancer Research Center Reducing cd33 expression to selectively protect therapeutic cells
JP2022501332A (en) 2018-09-19 2022-01-06 ジェネンテック， インコーポレイテッド How to treat and diagnose bladder cancer
KR20210089146A (en) 2018-09-19 2021-07-15 알파인 이뮨 사이언시즈, 인코포레이티드 Methods and uses of variant CD80 proteins and related constructs
ES2955032T3 (en) 2018-09-21 2023-11-28 Hoffmann La Roche Diagnostic methods for triple negative breast cancer
WO2020064702A1 (en) 2018-09-25 2020-04-02 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of antagonists of th17 cytokines for the treatment of bronchial remodeling in patients suffering from allergic asthma
CN112513088B (en) 2018-09-26 2023-05-16 江苏恒瑞医药股份有限公司 anti-OX 40 antibodies, antigen binding fragments thereof, and medical uses thereof
WO2020069398A1 (en) 2018-09-27 2020-04-02 Akrevia Therapeutics Inc. Masked cytokine polypeptides
WO2020070062A1 (en) 2018-10-01 2020-04-09 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of tim-3 inhibitors for the treatment of exacerbations in patients suffering from severe asthma
EP3861022A1 (en) 2018-10-04 2021-08-11 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical composition for the treatment of mucosal inflammatory diseases
JP2022512648A (en) 2018-10-09 2022-02-07 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル Use of αV-integrin (CD51) inhibitors for the treatment of myocardial fibrosis
WO2020076969A2 (en) 2018-10-10 2020-04-16 Tilos Therapeutics, Inc. Anti-lap antibody variants and uses thereof
UY38407A (en) 2018-10-15 2020-05-29 Novartis Ag TREM2 STABILIZING ANTIBODIES
WO2020081493A1 (en) 2018-10-16 2020-04-23 Molecular Templates, Inc. Pd-l1 binding proteins
WO2020081767A1 (en) 2018-10-18 2020-04-23 Genentech, Inc. Diagnostic and therapeutic methods for sarcomatoid kidney cancer
PE20211279A1 (en) 2018-10-23 2021-07-19 Dragonfly Therapeutics Inc HETERODIMERIC PROTEINS FUSED WITH FC
WO2020089437A1 (en) 2018-10-31 2020-05-07 Engmab Sàrl Combination therapy
US20210395390A1 (en) 2018-10-31 2021-12-23 Bayer Aktiengesellschaft Reversal agents for neutralizing the therapeutic activity of anti-fxia antibodies
WO2020089811A1 (en) 2018-10-31 2020-05-07 Novartis Ag Dc-sign antibody drug conjugates
JP2022506156A (en) 2018-11-05 2022-01-17 ジェネンテック， インコーポレイテッド Method for producing double-stranded protein in prokaryotic host cell
TW202039831A (en) 2018-11-05 2020-11-01 美商艾歐凡斯生物治療公司 Treatment of nsclc patients refractory for anti-pd-1 antibody
MX2021005751A (en) 2018-11-16 2021-10-01 Memorial Sloan Kettering Cancer Center Antibodies to mucin-16 and methods of use thereof.
TWI779253B (en) 2018-11-27 2022-10-01 大陸商信達生物製藥（蘇州）有限公司 ANTI-IL-23p19 ANTIBODY AND USE THEREOF
WO2020112781A1 (en) 2018-11-28 2020-06-04 Bristol-Myers Squibb Company Antibodies comprising modified heavy constant regions
JP2022510276A (en) 2018-11-30 2022-01-26 アルパイン イミューン サイエンシズ インコーポレイテッド CD86 variant immunomodulatory protein and its use
CA3121265A1 (en) 2018-12-05 2020-06-11 Genentech, Inc. Diagnostic methods and compositions for cancer immunotherapy
CA3121699A1 (en) 2018-12-05 2020-06-11 Morphosys Ag Multispecific antigen-binding molecules
WO2020117257A1 (en) 2018-12-06 2020-06-11 Genentech, Inc. Combination therapy of diffuse large b-cell lymphoma comprising an anti-cd79b immunoconjugates, an alkylating agent and an anti-cd20 antibody
WO2020118011A1 (en) 2018-12-06 2020-06-11 Alexion Pharmaceuticals, Inc. Anti-alk2 antibodies and uses thereof
EP3894427A1 (en) 2018-12-10 2021-10-20 Genentech, Inc. Photocrosslinking peptides for site specific conjugation to fc-containing proteins
CA3123512A1 (en) 2018-12-18 2020-06-25 Catapult Therapeutics B.V. The use of anti-ccr7 mabs for the prevention or treatment of graft-versus-host disease (gvhd)
TW202039554A (en) 2018-12-19 2020-11-01 瑞士商諾華公司 Anti-tnf-alpha antibodies
AR117327A1 (en) 2018-12-20 2021-07-28 23Andme Inc ANTI-CD96 ANTIBODIES AND METHODS OF USE OF THEM
JP2022514290A (en) 2018-12-20 2022-02-10 ジェネンテック， インコーポレイテッド Modified antibody FC and usage
SG11202106116QA (en) 2018-12-21 2021-07-29 Genentech Inc Methods of producing polypeptides using a cell line resistant to apoptosis
KR20210107721A (en) 2018-12-21 2021-09-01 23앤드미 인코포레이티드 Anti-IL-36 antibodies and methods of use thereof
US11130804B2 (en) 2018-12-21 2021-09-28 Hoffmann-La Roche Inc. Antibody that binds to VEGF and IL-1beta and methods of use
CN111349162A (en) 2018-12-21 2020-06-30 神州细胞工程有限公司 Humanized anti-PD-1 antibodies and uses thereof
CA3234463A1 (en) 2018-12-21 2019-12-18 Novartis Ag Antibodies to pmel17 and conjugates thereof
KR20210141447A (en) 2018-12-26 2021-11-23 실리오 디벨럽먼트, 인크. Anti-CTLA4 antibodies and methods of use thereof
EP3902564A4 (en) * 2018-12-27 2022-09-28 Kindred Biosciences, Inc. Igg fc variants for veterinary use
WO2020141145A1 (en) 2018-12-30 2020-07-09 F. Hoffmann-La Roche Ag Anti-rabbit cd19 antibodies and methods of use
MX2021008121A (en) 2019-01-03 2021-12-10 Inst Nat Sante Rech Med Methods and pharmaceutical compositions for enhancing cd8+ t cell-dependent immune responses in subjects suffering from cancer.
CN113597319A (en) 2019-01-04 2021-11-02 分解治疗有限责任公司 Treatment of xerosis with nuclease fusion proteins
MX2021008434A (en) 2019-01-14 2021-09-23 Genentech Inc Methods of treating cancer with a pd-1 axis binding antagonist and an rna vaccine.
WO2020148207A1 (en) 2019-01-14 2020-07-23 INSERM (Institut National de la Santé et de la Recherche Médicale) Human monoclonal antibodies binding to hla-a2
SG11202107538VA (en) 2019-01-16 2021-08-30 Compass Therapeutics Llc Formulations of antibodies that bind human cd137 and uses thereof
WO2020154293A1 (en) 2019-01-22 2020-07-30 Bristol-Myers Squibb Company Antibodies against il-7r alpha subunit and uses thereof
EP3914291A2 (en) 2019-01-22 2021-12-01 F. Hoffmann-La Roche AG Immunoglobulin a antibodies and methods of production and use
CA3124515A1 (en) 2019-01-23 2020-07-30 Genentech, Inc. Methods of producing multimeric proteins in eukaryotic host cells
WO2020154595A1 (en) 2019-01-24 2020-07-30 Massachusetts Institute Of Technology Nucleic acid nanostructure platform for antigen presentation and vaccine formulations formed therefrom
EP3915581A4 (en) 2019-01-24 2023-03-22 Chugai Seiyaku Kabushiki Kaisha Novel cancer antigens and antibodies of said antigens
GB201901197D0 (en) 2019-01-29 2019-03-20 Femtogenix Ltd G-A Crosslinking cytotoxic agents
MX2021010313A (en) 2019-02-27 2021-09-23 Genentech Inc Dosing for treatment with anti-tigit and anti-cd20 or anti-cd38 antibodies.
BR112021016875A2 (en) 2019-03-01 2022-01-04 Iovance Biotherapeutics Inc Process for expansion of peripheral blood lymphocytes
AU2020238811A1 (en) 2019-03-08 2021-07-22 Genentech, Inc. Methods for detecting and quantifying membrane-associated proteins on extracellular vesicles
US20220135682A1 (en) 2019-03-11 2022-05-05 Jounce Therapeutics, Inc. Anti-ICOS Antibodies for the Treatment of Cancer
CA3130446A1 (en) 2019-03-14 2020-09-17 Genentech, Inc. Treatment of cancer with her2xcd3 bispecific antibodies in combination with anti-her2 mab
EP3941940A1 (en) 2019-03-20 2022-01-26 Imcheck Therapeutics SAS Antibodies having specificity for btn2 and uses thereof
US20220175815A1 (en) 2019-04-03 2022-06-09 Orega Biotech Combination therapies based on pd1 and il-17b inhibitors
MA55529A (en) 2019-04-03 2022-02-09 Genzyme Corp REDUCED FRAGMENTATION ANTI-ALPHA BETA TCR BINDING POLYPEPTIDES
MX2021012160A (en) 2019-04-08 2022-01-06 Biogen Ma Inc Anti-integrin antibodies and uses thereof.
GB2589049C (en) 2019-04-11 2024-02-21 argenx BV Anti-IgE antibodies
US11136353B2 (en) 2019-04-15 2021-10-05 Qwixel Therapeutics Llc Fusion protein composition(s) comprising masked type I interferons (IFNA and IFNB) for use in the treatment of cancer and methods thereof
CA3136816A1 (en) 2019-04-17 2020-10-22 Alpine Immune Sciences, Inc. Methods and uses of variant icos ligand (icosl) fusion proteins
AU2020258480A1 (en) 2019-04-19 2021-10-21 Genentech, Inc. Anti-mertk antibodies and their methods of use
WO2020226986A2 (en) 2019-05-03 2020-11-12 Genentech, Inc. Methods of treating cancer with an anti-pd-l1 antibody
AU2020275415A1 (en) 2019-05-14 2021-11-25 Genentech, Inc. Methods of using anti-CD79B immunoconjugates to treat follicular lymphoma
AU2020279731A1 (en) 2019-05-20 2022-01-06 Novartis Ag Antibody drug conjugates having linkers comprising hydrophilic groups
US20230092679A1 (en) 2019-05-20 2023-03-23 Novartis Ag Mcl-1 inhibitor antibody-drug conjugates and methods of use
WO2020234399A1 (en) 2019-05-20 2020-11-26 INSERM (Institut National de la Santé et de la Recherche Médicale) Novel anti-cd25 antibodies
EP3972993A1 (en) 2019-05-21 2022-03-30 Novartis AG Variant cd58 domains and uses thereof
US20220396631A1 (en) 2019-05-21 2022-12-15 Lu HUANG Trispecific binding molecules against bcma and uses thereof
JOP20210309A1 (en) 2019-05-21 2023-01-30 Novartis Ag Cd19 binding molecules and uses thereof
AU2020278907A1 (en) 2019-05-23 2022-01-20 Ac Immune Sa Anti-TDP-43 binding molecules and uses thereof
MX2021014756A (en) 2019-06-07 2022-01-18 Argenx Bvba PHARMACEUTICAL FORMULATIONS OF FcRn INHIBITORS SUITABLE FOR SUBCUTANEOUS ADMINISTRATION.
AU2020291300A1 (en) 2019-06-10 2022-01-06 Chugai Seiyaku Kabushiki Kaisha Anti-T cell antigen-binding molecule to be used in combination with cytokine inhibitor
CR20210607A (en) 2019-06-12 2022-01-21 Novartis Ag Natriuretic peptide receptor 1 antibodies and methods of use
EP3986918A1 (en) 2019-06-18 2022-04-27 Bayer Aktiengesellschaft Adrenomedullin-analogues for long-term stabilization and their use
US11905532B2 (en) 2019-06-25 2024-02-20 Massachusetts Institute Of Technology Compositions and methods for molecular memory storage and retrieval
CA3139250A1 (en) 2019-07-10 2021-01-14 Naoki Kimura Claudin-6 binding molecules and uses thereof
WO2021005232A1 (en) 2019-07-11 2021-01-14 Umc Utrecht Holding B.V. Intranasal administration of neutralising antiviral antibodies
AR119382A1 (en) 2019-07-12 2021-12-15 Hoffmann La Roche PRE-TARGETING ANTIBODIES AND METHODS OF USE
CN114341187A (en) 2019-07-12 2022-04-12 中外制药株式会社 Anti-mutant FGFR3 antibodies and uses thereof
WO2021011678A1 (en) 2019-07-15 2021-01-21 Bristol-Myers Squibb Company Anti-trem-1 antibodies and uses thereof
JP2022540904A (en) 2019-07-15 2022-09-20 ブリストル－マイヤーズ スクイブ カンパニー Antibodies against human TREM-1 and uses thereof
WO2021009263A1 (en) 2019-07-16 2021-01-21 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies having specificity for cd38 and uses thereof
BR112022001017A2 (en) 2019-07-19 2022-04-12 Sinocelltech Ltd Humanized anti-vegf monoclonal antibody
WO2021013065A1 (en) 2019-07-19 2021-01-28 神州细胞工程有限公司 Humanized anti-vegf fab antibody fragment and use thereof
CN114144436A (en) 2019-07-24 2022-03-04 H.隆德贝克有限公司 anti-mGluR 5 antibodies and uses thereof
US20220291203A1 (en) 2019-07-25 2022-09-15 Immunowake Inc. Methods of measuring cell-mediated killing by effectors
CN112300279A (en) 2019-07-26 2021-02-02 上海复宏汉霖生物技术股份有限公司 Methods and compositions directed to anti-CD 73 antibodies and variants
KR20220071179A (en) 2019-07-26 2022-05-31 사이노셀테크 엘티디. Humanized anti-IL17A antibodies and uses thereof
MX2022001146A (en) 2019-07-29 2022-03-17 Compugen Ltd Anti-pvrig antibodies formulations and uses thereof.
TW202120124A (en) 2019-07-31 2021-06-01 瑞士商赫孚孟拉羅股份公司 Dosage and administration regimen for the treatment or prevention of c5-related diseases by the use of the anti-c5 antibody crovalimab
MX2022001154A (en) 2019-07-31 2022-02-22 Hoffmann La Roche Dosage and administration regimen for the treatment or prevention of c5-related diseases by the use of the anti-c5 antibody crovalimab.
KR102509648B1 (en) 2019-08-06 2023-03-15 아프리노이아 테라퓨틱스 리미티드 Antibodies that bind to pathological Tau species and uses thereof
MX2022001882A (en) 2019-08-12 2022-05-30 Aptevo Res & Development Llc 4-1bb and ox40 binding proteins and related compositions and methods, antibodies against 4-1bb, antibodies against ox40.
KR20220053007A (en) 2019-08-30 2022-04-28 아게누스 인코포레이티드 Anti-CD96 antibodies and methods of use thereof
CN114341183B (en) 2019-09-04 2024-04-16 Inserm(法国国家健康医学研究院) IL20-RB specific antibodies and their use for treating acute exacerbations of chronic obstructive pulmonary disease
CN114340675A (en) 2019-09-12 2022-04-12 豪夫迈·罗氏有限公司 Compositions and methods for treating lupus nephritis
AU2020351122A1 (en) 2019-09-16 2022-02-17 Surface Oncology, Inc. Anti-CD39 antibody compositions and methods
WO2021055816A1 (en) 2019-09-18 2021-03-25 Molecular Templates, Inc. Pd-l1 binding molecules comprising shiga toxin a subunit scaffolds
TW202124446A (en) 2019-09-18 2021-07-01 瑞士商諾華公司 Combination therapies with entpd2 antibodies
PE20221906A1 (en) 2019-09-18 2022-12-23 Genentech Inc ANTI-KLK7 ANTIBODIES, ANTI-KLK5 ANTIBODIES, ANTI-KLK5/KLK7 MULTISPECIFIC ANTIBODIES AND METHODS OF USE
US20220348651A1 (en) 2019-09-18 2022-11-03 Novartis Ag Entpd2 antibodies, combination therapies, and methods of using the antibodies and combination therapies
MX2022003204A (en) 2019-09-19 2022-04-18 Bristol Myers Squibb Co Antibodies binding to vista at acidic ph.
CR20220149A (en) 2019-09-20 2022-05-23 Genentech Inc Dosing for anti-tryptase antibodies
BR112022004302A2 (en) 2019-09-25 2022-06-21 Surface Oncology Inc Anti-il-27 antibodies and uses thereof
WO2021062323A1 (en) 2019-09-26 2021-04-01 Stcube & Co. Antibodies specific to glycosylated ctla-4 and methods of use thereof
JP2022548978A (en) 2019-09-27 2022-11-22 ジェネンテック， インコーポレイテッド Dosing for Treatment with Drugs Anti-TIGIT and Anti-PD-L1 Antagonist Antibodies
WO2021059075A1 (en) 2019-09-27 2021-04-01 Janssen Biotech, Inc. Anti-ceacam antibodies and uses thereof
EP4038182A1 (en) 2019-09-30 2022-08-10 Bioverativ Therapeutics Inc. Lentiviral vector formulations
EP3800201A1 (en) 2019-10-01 2021-04-07 INSERM (Institut National de la Santé et de la Recherche Médicale) Cd28h stimulation enhances nk cell killing activities
US20240058466A1 (en) 2019-10-04 2024-02-22 TAE Life Sciences Antibody Compositions Comprising Fc Mutations and Site-Specific Conjugation Properties For Use In Treating Cancer, Immunological Disorders, and Methods Thereof
WO2021072277A1 (en) 2019-10-09 2021-04-15 Stcube & Co. Antibodies specific to glycosylated lag3 and methods of use thereof
JP7413519B2 (en) 2019-10-18 2024-01-15 ジェネンテック， インコーポレイテッド Methods of using anti-CD79B immunoconjugates to treat diffuse large B-cell lymphoma
US11459389B2 (en) 2019-10-24 2022-10-04 Massachusetts Institute Of Technology Monoclonal antibodies that bind human CD161
IL292419A (en) 2019-10-24 2022-06-01 Prometheus Biosciences Inc Humanized antibodies to tnf-like ligand 1a (tl1a) and uses thereof
WO2021086948A1 (en) 2019-10-28 2021-05-06 Georgia Tech Research Corporation Mrna-encoded antibodies for contraception
WO2021086953A1 (en) 2019-10-28 2021-05-06 Georgia Tech Research Corporation Compositions and methods for prophylaxis of hiv
MX2022005132A (en) 2019-11-04 2022-08-15 Compugen Ltd Combination therapy with anti-pvrig antibodies formulations and anti-pd-1 antibodies.
US20220378909A1 (en) 2019-11-05 2022-12-01 Jounce Therapeutics, Inc. Methods of Treating Cancer with Anti-PD-1 Antibodies
IL292458A (en) 2019-11-06 2022-06-01 Genentech Inc Diagnostic and therapeutic methods for treatment of hematologic cancers
US20230040928A1 (en) 2019-12-09 2023-02-09 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies having specificity to her4 and uses thereof
AU2020403145A1 (en) 2019-12-13 2022-07-07 Genentech, Inc. Anti-Ly6G6D antibodies and methods of use
JP2023506956A (en) 2019-12-20 2023-02-20 エフ・ホフマン－ラ・ロシュ・アクチェンゲゼルシャフト IL-37 fusion proteins and uses thereof
CN113045655A (en) 2019-12-27 2021-06-29 高诚生物医药(香港)有限公司 anti-OX 40 antibodies and uses thereof
AU2019479791A1 (en) 2019-12-27 2022-07-14 Chugai Seiyaku Kabushiki Kaisha Anti-CTLA-4 antibody and use thereof
AU2021205877A1 (en) 2020-01-06 2022-06-09 Vaccinex, Inc. Anti-CCR8 antibodies and uses thereof
US20210236596A1 (en) 2020-01-08 2021-08-05 argenx BV Methods for treating pemphigus disorders
CN110818795B (en) 2020-01-10 2020-04-24 上海复宏汉霖生物技术股份有限公司 anti-TIGIT antibodies and methods of use
KR20220131293A (en) 2020-01-24 2022-09-27 화이자 인코포레이티드 Anti-E-Selectin Antibodies, Compositions and Methods of Use
WO2021194481A1 (en) 2020-03-24 2021-09-30 Genentech, Inc. Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies
WO2022050954A1 (en) 2020-09-04 2022-03-10 Genentech, Inc. Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies
WO2021155295A1 (en) 2020-01-31 2021-08-05 The Cleveland Clinic Foundation Anti-müllerian hormone receptor 2 antibodies and methods of use
EP4096708A1 (en) 2020-01-31 2022-12-07 Genentech, Inc. Methods of inducing neoepitope-specific t cells with a pd-1 axis binding antagonist and an rna vaccine
US20230087600A1 (en) 2020-02-06 2023-03-23 Bristol-Myers Squibb Company Il-10 and uses thereof
TW202144395A (en) 2020-02-12 2021-12-01 日商中外製藥股份有限公司 Anti-CD137 antigen-binding molecule for use in cancer treatment
WO2021163064A2 (en) 2020-02-14 2021-08-19 Jounce Therapeutics, Inc. Antibodies and fusion proteins that bind to ccr8 and uses thereof
CA3169910A1 (en) 2020-02-28 2021-09-02 Shanghai Henlius Biotech, Inc. Anti-cd137 constructs, multispecific antibody and uses thereof
CN115066440A (en) 2020-02-28 2022-09-16 上海复宏汉霖生物技术股份有限公司 anti-CD 137 constructs and uses thereof
EP4110810A1 (en) 2020-02-28 2023-01-04 Orega Biotech Combination therapies based on ctla4 and il-17b inhibitors
JP2023516952A (en) 2020-02-28 2023-04-21 ジェンザイム・コーポレーション Modified binding polypeptides for optimized drug conjugation
WO2021175954A1 (en) 2020-03-04 2021-09-10 Imcheck Therapeutics Sas Antibodies having specificity for btnl8 and uses thereof
MX2022011178A (en) 2020-03-10 2023-01-04 Massachusetts Inst Technology COMPOSITIONS AND METHODS FOR IMMUNOTHERAPY OF NPM1c-POSITIVE CANCER.
IL296256A (en) 2020-03-13 2022-11-01 Genentech Inc Anti-interleukin-33 antibodies and uses thereof
CN117510630A (en) 2020-03-19 2024-02-06 基因泰克公司 Isotype selective anti-TGF-beta antibodies and methods of use
CN115666589A (en) 2020-03-19 2023-01-31 艾维迪提生物科学公司 Compositions and methods for treating facioscapulohumeral muscular dystrophy
US11365239B2 (en) 2020-03-20 2022-06-21 Tsb Therapeutics (Beijing) Co., Ltd. Anti-SARS-COV-2 antibodies and uses thereof
WO2021194913A1 (en) 2020-03-24 2021-09-30 Genentech, Inc. Tie2-binding agents and methods of use
CN111122879B (en) * 2020-03-26 2020-07-14 珠海丽珠试剂股份有限公司 Coating liquid of novel coronavirus recombinant antigen, pretreatment method, application and product
EP4127153A2 (en) 2020-03-26 2023-02-08 Genentech, Inc. Modified mammalian cells having reduced host cell proteins
CN116249549A (en) 2020-03-27 2023-06-09 诺华股份有限公司 Bispecific combination therapies for the treatment of proliferative diseases and autoimmune disorders
CN115397850A (en) 2020-03-30 2022-11-25 豪夫迈·罗氏有限公司 Antibodies that bind to VEGF and PDGF-B and methods of use thereof
CA3170570A1 (en) 2020-04-01 2021-10-07 James J. KOBIE Monoclonal antibodies against the hemagglutinin (ha) and neuraminidase (na) of influenza h3n2 viruses
WO2021202959A1 (en) 2020-04-03 2021-10-07 Genentech, Inc. Therapeutic and diagnostic methods for cancer
US20230132275A1 (en) 2020-04-08 2023-04-27 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of cdon inhibitors for the treatment of endothelial dysfunction
TW202204622A (en) 2020-04-09 2022-02-01 大陸商蘇州艾博生物科技有限公司 Nucleic acid vaccines for coronavirus
WO2021207449A1 (en) 2020-04-09 2021-10-14 Merck Sharp & Dohme Corp. Affinity matured anti-lap antibodies and uses thereof
WO2021207662A1 (en) 2020-04-10 2021-10-14 Genentech, Inc. Use of il-22fc for the treatment or prevention of pneumonia, acute respiratory distress syndrome, or cytokine release syndrome
IL297313A (en) 2020-04-20 2022-12-01 Jounce Therapeutics Inc Compositions and methods for vaccination and the treatment of infectious diseases
WO2021214277A1 (en) 2020-04-24 2021-10-28 F. Hoffmann-La Roche Ag Enzyme and pathway modulation with sulfhydryl compounds and their derivatives
IL297541A (en) 2020-04-24 2022-12-01 Genentech Inc Methods of using anti-cd79b immunoconjugates
SG11202112792WA (en) 2020-04-28 2021-12-30 Univ Rockefeller Neutralizing anti-sars-cov-2 antibodies and methods of use thereof
WO2021222167A1 (en) 2020-04-28 2021-11-04 Genentech, Inc. Methods and compositions for non-small cell lung cancer immunotherapy
US20230181756A1 (en) 2020-04-30 2023-06-15 Novartis Ag Ccr7 antibody drug conjugates for treating cancer
WO2021219871A2 (en) 2020-04-30 2021-11-04 Aduro Biotech Holdings, Europe B.V. Anti-cd103 antibodies
WO2021226551A1 (en) 2020-05-08 2021-11-11 Alpine Immune Sciences, Inc. April and baff inhibitory immunomodulatory proteins and methods of use thereof
JP2023525053A (en) 2020-05-12 2023-06-14 インサーム（インスティテュ ナシオナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシェ メディカル） A new method to treat cutaneous T-cell lymphoma and TFH-derived lymphoma
WO2021231732A1 (en) 2020-05-15 2021-11-18 Bristol-Myers Squibb Company Antibodies to garp
WO2021233834A1 (en) 2020-05-17 2021-11-25 Astrazeneca Uk Limited Sars-cov-2 antibodies and methods of selecting and using the same
GB2595299B (en) 2020-05-21 2022-08-03 Mabsolve Ltd Modified immunoglobulin FC regions
EP4157881A1 (en) 2020-05-27 2023-04-05 Staidson (Beijing) Biopharmaceuticals Co., Ltd. Antibodies specifically recognizing nerve growth factor and uses thereof
AU2021281417A1 (en) 2020-05-29 2022-12-08 23Andme, Inc. Anti-CD200R1 antibodies and methods of use thereof
IL298735A (en) 2020-06-02 2023-02-01 Dynamicure Biotechnology Llc Anti-cd93 constructs and uses thereof
CN116529260A (en) 2020-06-02 2023-08-01 当康生物技术有限责任公司 anti-CD 93 constructs and uses thereof
AR122569A1 (en) 2020-06-08 2022-09-21 Hoffmann La Roche ANTI-HBV ANTIBODIES AND METHODS OF USE
WO2021252977A1 (en) 2020-06-12 2021-12-16 Genentech, Inc. Methods and compositions for cancer immunotherapy
KR20230025691A (en) 2020-06-16 2023-02-22 제넨테크, 인크. Methods and compositions for treating triple negative breast cancer
KR20230024368A (en) 2020-06-18 2023-02-20 제넨테크, 인크. Treatment with anti-TIGIT antibodies and PD-1 axis binding antagonists
US20230235056A1 (en) 2020-06-19 2023-07-27 Chugai Seiyaku Kabushiki Kaisha Anti-t cell antigen-binding molecule for use in combination with angiogenesis inhibitor
EP4183802A1 (en) 2020-06-22 2023-05-24 Innovent Biologics (Suzhou) Co., Ltd. Anti-ang-2 antibody and use thereof
WO2021259880A1 (en) 2020-06-22 2021-12-30 Almirall, S.A. Anti-il-36 antibodies and methods of use thereof
US20230312741A1 (en) 2020-06-22 2023-10-05 Innovent Biologics (Suzhou) Co., Ltd. Anti-cd73 antibody and uses thereof
JP2023533217A (en) 2020-06-24 2023-08-02 ジェネンテック， インコーポレイテッド Apoptosis resistant cell line
JP2023528535A (en) 2020-06-25 2023-07-04 メルク・シャープ・アンド・ドーム・エルエルシー High Affinity Antibodies Targeting Tau Phosphorylated at Serine 413
CA3165342A1 (en) 2020-06-29 2022-01-06 James Arthur Posada Treatment of sjogren's syndrome with nuclease fusion proteins
MX2023000339A (en) 2020-07-10 2023-02-09 Hoffmann La Roche Antibodies which bind to cancer cells and target radionuclides to said cells.
PE20231300A1 (en) 2020-07-17 2023-08-24 Genentech Inc ANTI-NOTCH2 ANTIBODIES AND METHODS OF USE
GB2597532A (en) 2020-07-28 2022-02-02 Femtogenix Ltd Cytotoxic compounds
CA3190328A1 (en) 2020-07-29 2022-02-03 Dynamicure Biotechnology Llc Anti-cd93 constructs and uses thereof
JP2023536602A (en) 2020-08-03 2023-08-28 ジェネンテック， インコーポレイテッド Diagnostic and therapeutic methods for lymphoma
JP2023536904A (en) 2020-08-06 2023-08-30 バイオベラティブ・ユーエスエイ・インコーポレイテッド Inflammatory Cytokines and Fatigue in Subjects With Complement-Mediated Disease
WO2022034044A1 (en) 2020-08-10 2022-02-17 Astrazeneca Uk Limited Sars-cov-2 antibodies for treatment and prevention of covid-19
EP4204448A2 (en) 2020-08-27 2023-07-05 cureab GmbH Anti-golph2 antibodies for macrophage and dendritic cell differentiation
CA3192344A1 (en) 2020-08-28 2022-03-03 Genentech, Inc. Crispr/cas9 multiplex knockout of host cell proteins
KR20230061458A (en) 2020-09-04 2023-05-08 에프. 호프만-라 로슈 아게 Antibodies that bind to VEGF-A and ANG2 and methods of use
WO2022053715A1 (en) 2020-09-14 2022-03-17 Ichnos Sciences SA Antibodies that bind to il1rap and uses thereof
CN116406291A (en) 2020-10-05 2023-07-07 基因泰克公司 Administration of treatment with anti-FCRH 5/anti-CD 3 bispecific antibodies
WO2022076606A1 (en) 2020-10-06 2022-04-14 Iovance Biotherapeutics, Inc. Treatment of nsclc patients with tumor infiltrating lymphocyte therapies
JP2023546359A (en) 2020-10-06 2023-11-02 アイオバンス バイオセラピューティクス，インコーポレイテッド Treatment of NSCLC patients with tumor-infiltrating lymphocyte therapy
IL302029A (en) 2020-10-14 2023-06-01 Denali Therapeutics Inc Fusion proteins comprising sulfoglucosamine sulfohydrolase enzymes and methods thereof
AR123855A1 (en) 2020-10-20 2023-01-18 Genentech Inc PEG-CONJUGATED ANTI-MERTK ANTIBODIES AND METHODS OF USE
WO2022090801A2 (en) 2020-10-26 2022-05-05 Compugen Ltd. Pvrl2 and/or pvrig as biomarkers for treatment
WO2022093981A1 (en) 2020-10-28 2022-05-05 Genentech, Inc. Combination therapy comprising ptpn22 inhibitors and pd-l1 binding antagonists
AU2021374594A1 (en) 2020-11-04 2023-06-01 Genentech, Inc. Dosing for treatment with anti-cd20/anti-cd3 bispecific antibodies and anti-cd79b antibody drug conjugates
WO2022098870A1 (en) 2020-11-04 2022-05-12 The Rockefeller University Neutralizing anti-sars-cov-2 antibodies
CA3196076A1 (en) 2020-11-04 2022-05-12 Chi-Chung Li Subcutaneous dosing of anti-cd20/anti-cd3 bispecific antibodies
CA3196539A1 (en) 2020-11-04 2022-05-12 Chi-Chung Li Dosing for treatment with anti-cd20/anti-cd3 bispecific antibodies
WO2022096547A1 (en) 2020-11-05 2022-05-12 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of il-6 inhibitors for the treatment of acute chest syndrome in patients suffering from sickle cell disease
WO2022097060A1 (en) 2020-11-06 2022-05-12 Novartis Ag Cd19 binding molecules and uses thereof
CN116472288A (en) 2020-11-06 2023-07-21 诺华股份有限公司 Antibody Fc variants
MX2023005234A (en) 2020-11-06 2023-05-18 Novartis Ag Anti-cd19 agent and b cell targeting agent combination therapy for treating b cell malignancies.
WO2022105847A1 (en) 2020-11-19 2022-05-27 Staidson (Beijing) Biopharmaceuticals Co. , Ltd. Long-acting nerve growth factor polypeptides and uses thereof
JP2024504547A (en) 2020-11-20 2024-02-01 インセルム（インスティチュート ナショナル デ ラ サンテ エ デ ラ リシェルシェ メディカル） Anti-CD25 antibody
AR124123A1 (en) 2020-11-20 2023-02-15 Inst Nat Sante Rech Med ANTI-CD25 ANTIBODIES
EP4251648A2 (en) 2020-11-24 2023-10-04 Novartis AG Anti-cd48 antibodies, antibody drug conjugates, and uses thereof
JP2023553808A (en) 2020-11-24 2023-12-26 ノバルティス アーゲー MCL-1 inhibitor antibody-drug conjugates and methods of use
US20240042024A1 (en) 2020-11-27 2024-02-08 David A. Horwitz Methods and compositions for treatment of immune-mediated diseases
EP4255574A1 (en) 2020-12-01 2023-10-11 Aptevo Research and Development LLC Heterodimeric psma and cd3-binding bispecific antibodies
TW202241468A (en) 2020-12-11 2022-11-01 美商艾歐凡斯生物治療公司 Treatment of cancer patients with tumor infiltrating lymphocyte therapies in combination with braf inhibitors and/or mek inhibitors
WO2022130182A1 (en) 2020-12-14 2022-06-23 Novartis Ag Reversal binding agents for anti-natriuretic peptide receptor 1 (npr1) antibodies and uses thereof
EP4262827A1 (en) 2020-12-17 2023-10-25 Iovance Biotherapeutics, Inc. Treatment of cancers with tumor infiltrating lymphocytes
JP2023554395A (en) 2020-12-17 2023-12-27 アイオバンス バイオセラピューティクス，インコーポレイテッド Treatment with tumor-infiltrating lymphocyte therapy in combination with CTLA-4 and PD-1 inhibitors
KR20230124959A (en) 2020-12-23 2023-08-28 이노벤트 바이오로직스 (쑤저우) 컴퍼니, 리미티드 Anti-B7-H3 Antibodies and Uses Thereof
WO2022140797A1 (en) 2020-12-23 2022-06-30 Immunowake Inc. Immunocytokines and uses thereof
JP2024501845A (en) 2020-12-31 2024-01-16 アイオバンス バイオセラピューティクス，インコーポレイテッド Devices and processes for automated production of tumor-infiltrating lymphocytes
EP4277705A1 (en) 2021-01-12 2023-11-22 F. Hoffmann-La Roche AG Split antibodies which bind to cancer cells and target radionuclides to said cells
WO2022153212A1 (en) 2021-01-13 2022-07-21 Axon Neuroscience Se Antibodies neutralizing sars-cov-2
CA3204291A1 (en) 2021-01-13 2022-07-21 F. Hoffmann-La Roche Ag Combination therapy
EP4277926A1 (en) 2021-01-15 2023-11-22 The Rockefeller University Neutralizing anti-sars-cov-2 antibodies
US20240082397A1 (en) 2021-01-28 2024-03-14 Compugen Ltd. Anti-pvrig antibodies formulations and uses thereof
US20240076373A1 (en) 2021-01-28 2024-03-07 Compugen Ltd. Combination therapy with anti-pvrig antibodies formulations and anti-pd-1 antibodies
EP4284919A1 (en) 2021-01-29 2023-12-06 Iovance Biotherapeutics, Inc. Methods of making modified tumor infiltrating lymphocytes and their use in adoptive cell therapy
AU2022218137A1 (en) 2021-02-03 2023-08-24 Mozart Therapeutics, Inc. Binding agents and methods of using the same
WO2022169872A1 (en) 2021-02-03 2022-08-11 Genentech, Inc. Multispecific binding protein degrader platform and methods of use
EP4289865A1 (en) 2021-02-04 2023-12-13 Innovent Biologics (Suzhou) Co., Ltd. Anti-tnfr2 antibody and use thereof
AU2022224561A1 (en) 2021-02-17 2023-09-28 Dr. Falk Pharma Gmbh Anti-cd30l antibodies and uses thereof
CA3209364A1 (en) 2021-03-01 2022-09-09 Jennifer O'neil Combination of masked ctla4 and pd1/pdl1 antibodies for treating cancer
WO2022187270A1 (en) 2021-03-01 2022-09-09 Xilio Development, Inc. Combination of ctla4 and pd1/pdl1 antibodies for treating cancer
CA3210755A1 (en) 2021-03-05 2022-09-09 Kenneth ONIMUS Tumor storage and cell culture compositions
JP2024509191A (en) 2021-03-05 2024-02-29 ダイナミキュア バイオテクノロジー エルエルシー Anti-VISTA constructs and their uses
EP4294927A2 (en) 2021-03-10 2023-12-27 Immunowake Inc. Immunomodulatory molecules and uses thereof
WO2022192647A1 (en) 2021-03-12 2022-09-15 Genentech, Inc. Anti-klk7 antibodies, anti-klk5 antibodies, multispecific anti-klk5/klk7 antibodies, and methods of use
WO2022198192A1 (en) 2021-03-15 2022-09-22 Genentech, Inc. Compositions and methods of treating lupus nephritis
CA3213295A1 (en) 2021-03-17 2022-09-22 Molecular Templates, Inc. Pd-l1 binding proteins comprising shiga toxin a subunit scaffolds and cd8+ t cell antigens
CA3213636A1 (en) 2021-03-18 2022-09-22 Seagen Inc. Selective drug release from internalized conjugates of biologically active compounds
WO2022198141A1 (en) 2021-03-19 2022-09-22 Iovance Biotherapeutics, Inc. Methods for tumor infiltrating lymphocyte (til) expansion related to cd39/cd69 selection and gene knockout in tils
WO2022197877A1 (en) 2021-03-19 2022-09-22 Genentech, Inc. Methods and compositions for time delayed bio-orthogonal release of cytotoxic agents
EP4313122A1 (en) 2021-03-23 2024-02-07 Iovance Biotherapeutics, Inc. Cish gene editing of tumor infiltrating lymphocytes and uses of same in immunotherapy
EP4314049A1 (en) 2021-03-25 2024-02-07 Dynamicure Biotechnology LLC Anti-igfbp7 constructs and uses thereof
EP4314253A2 (en) 2021-03-25 2024-02-07 Iovance Biotherapeutics, Inc. Methods and compositions for t-cell coculture potency assays and use with cell therapy products
JP2024513837A (en) 2021-03-31 2024-03-27 バイオベラティブ・ユーエスエイ・インコーポレイテッド Reducing surgery-related hemolysis in patients with cold agglutinin disease
EP4314068A1 (en) 2021-04-02 2024-02-07 The Regents Of The University Of California Antibodies against cleaved cdcp1 and uses thereof
CN117279664A (en) 2021-04-10 2023-12-22 普方生物制药美国公司 FOLR1 binding agents, conjugates thereof, and methods of use thereof
CA3216880A1 (en) 2021-04-16 2022-10-20 Novartis Ag Antibody drug conjugates and methods for making thereof
BR112023021665A2 (en) 2021-04-19 2023-12-19 Iovance Biotherapeutics Inc METHOD FOR TREATING A CANCER, AND, COMPOSITION
JP2024514222A (en) 2021-04-19 2024-03-28 ジェネンテック， インコーポレイテッド Modified Mammalian Cells
EP4326768A1 (en) 2021-04-23 2024-02-28 Profoundbio Us Co. Anti-cd70 antibodies, conjugates thereof and methods of using the same
EP4334343A2 (en) 2021-05-06 2024-03-13 The Rockefeller University Neutralizing anti-sars- cov-2 antibodies and methods of use thereof
IL308336A (en) 2021-05-07 2024-01-01 Alpine Immune Sciences Inc Methods of dosing and treatment with a taci-fc fusion immunomodulatory protein
CA3215856A1 (en) 2021-05-07 2022-11-10 Alison O'neill Anti-il-27 antibodies and uses thereof
CA3218170A1 (en) 2021-05-12 2022-11-17 Jamie Harue HIRATA Methods of using anti-cd79b immunoconjugates to treat diffuse large b-cell lymphoma
WO2022241082A1 (en) 2021-05-14 2022-11-17 Genentech, Inc. Agonists of trem2
WO2022245754A1 (en) 2021-05-17 2022-11-24 Iovance Biotherapeutics, Inc. Pd-1 gene-edited tumor infiltrating lymphocytes and uses of same in immunotherapy
WO2022246259A1 (en) 2021-05-21 2022-11-24 Genentech, Inc. Modified cells for the production of a recombinant product of interest
TW202306994A (en) 2021-06-04 2023-02-16 日商中外製藥股份有限公司 Anti-ddr2 antibodies and uses thereof
AU2022293999A1 (en) 2021-06-14 2023-11-30 argenx BV Anti-il-9 antibodies and methods of use thereof
EP4355785A1 (en) 2021-06-17 2024-04-24 Amberstone Biosciences, Inc. Anti-cd3 constructs and uses thereof
IL308959A (en) 2021-06-18 2024-01-01 Nammi Therapeutics Inc Fusion protein composition(s) comprising masked type i interferons (ifnα and ifnβ) for use in the treatment of cancer and methods thereof
IL308134A (en) 2021-06-22 2023-12-01 Novartis Ag Bispecific antibodies for use in treatment of hidradenitis suppurativa
EP4361176A1 (en) 2021-06-25 2024-05-01 Chugai Seiyaku Kabushiki Kaisha Use of anti-ctla-4 antibody
EP4361273A1 (en) 2021-06-25 2024-05-01 Chugai Seiyaku Kabushiki Kaisha Anti-ctla-4 antibody
WO2023275621A1 (en) 2021-07-01 2023-01-05 Compugen Ltd. Anti-tigit and anti-pvrig in monotherapy and combination treatments
TW202320857A (en) 2021-07-06 2023-06-01 美商普方生物製藥美國公司 Linkers, drug linkers and conjugates thereof and methods of using the same
TW202317633A (en) 2021-07-08 2023-05-01 美商舒泰神（加州）生物科技有限公司 Antibodies specifically recognizing tnfr2 and uses thereof
WO2023285362A1 (en) 2021-07-12 2023-01-19 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of il-36 inhibitors for the treatment of netherton syndrome
WO2023288182A1 (en) 2021-07-12 2023-01-19 Genentech, Inc. Structures for reducing antibody-lipase binding
IL309856A (en) 2021-07-14 2024-02-01 Genentech Inc Anti-c-c motif chemokine receptor 8 (ccr8) antibodies and methods of use
WO2023284714A1 (en) 2021-07-14 2023-01-19 舒泰神（北京）生物制药股份有限公司 Antibody that specifically recognizes cd40 and application thereof
WO2023001884A1 (en) 2021-07-22 2023-01-26 F. Hoffmann-La Roche Ag Heterodimeric fc domain antibodies
WO2023004074A2 (en) 2021-07-22 2023-01-26 Iovance Biotherapeutics, Inc. Method for cryopreservation of solid tumor fragments
WO2023004386A1 (en) 2021-07-22 2023-01-26 Genentech, Inc. Brain targeting compositions and methods of use thereof
WO2023010060A2 (en) 2021-07-27 2023-02-02 Novab, Inc. Engineered vlrb antibodies with immune effector functions
CA3226942A1 (en) 2021-07-28 2023-02-02 Iovance Biotherapeutics, Inc. Treatment of cancer patients with tumor infiltrating lymphocyte therapies in combination with kras inhibitors
AU2022324406A1 (en) 2021-08-02 2024-03-14 Innovent Biologics (Suzhou) Co., Ltd. Anti-cd79b×cd3 bispecific antibody and use thereof
CN117794953A (en) 2021-08-03 2024-03-29 豪夫迈·罗氏有限公司 Bispecific antibodies and methods of use
WO2023019092A1 (en) 2021-08-07 2023-02-16 Genentech, Inc. Methods of using anti-cd79b immunoconjugates to treat diffuse large b-cell lymphoma
WO2023019239A1 (en) 2021-08-13 2023-02-16 Genentech, Inc. Dosing for anti-tryptase antibodies
WO2023021055A1 (en) 2021-08-19 2023-02-23 F. Hoffmann-La Roche Ag Multivalent anti-variant fc-region antibodies and methods of use
AU2022332303A1 (en) 2021-08-27 2024-02-01 Genentech, Inc. Methods of treating tau pathologies
TW202325727A (en) 2021-08-30 2023-07-01 美商建南德克公司 Anti-polyubiquitin multispecific antibodies
WO2023039488A1 (en) 2021-09-09 2023-03-16 Iovance Biotherapeutics, Inc. Processes for generating til products using pd-1 talen knockdown
CA3232700A1 (en) 2021-09-24 2023-03-30 Rafael CUBAS Expansion processes and agents for tumor infiltrating lymphocytes
WO2023056403A1 (en) 2021-09-30 2023-04-06 Genentech, Inc. Methods for treatment of hematologic cancers using anti-tigit antibodies, anti-cd38 antibodies, and pd-1 axis binding antagonists
WO2023052541A1 (en) 2021-09-30 2023-04-06 Imcheck Therapeutics Combination of an anti-btn3a activating antibody and an il-2 agonist for use in therapy
WO2023056069A1 (en) 2021-09-30 2023-04-06 Angiex, Inc. Degrader-antibody conjugates and methods of using same
CA3232806A1 (en) 2021-09-30 2023-04-06 Seagen Inc. B7-h4 antibody-drug conjugates for the treatment of cancer
CN116064598B (en) 2021-10-08 2024-03-12 苏州艾博生物科技有限公司 Nucleic acid vaccine for coronavirus
WO2023064958A1 (en) 2021-10-15 2023-04-20 Compugen Ltd. Combination therapy with anti-pvrig antibodies formulations, anti-tigit antibodies, and anti-pd-1 antibodies
WO2023076876A1 (en) 2021-10-26 2023-05-04 Mozart Therapeutics, Inc. Modulation of immune responses to viral vectors
TW202331735A (en) 2021-10-27 2023-08-01 美商艾歐凡斯生物治療公司 Systems and methods for coordinating manufacturing of cells for patient-specific immunotherapy
TW202342095A (en) 2021-11-05 2023-11-01 英商阿斯特捷利康英國股份有限公司 Composition for treatment and prevention of covid-19
WO2023081818A1 (en) 2021-11-05 2023-05-11 American Diagnostics & Therapy, Llc (Adxrx) Monoclonal antibodies against carcinoembryonic antigens, and their uses
WO2023086807A1 (en) 2021-11-10 2023-05-19 Genentech, Inc. Anti-interleukin-33 antibodies and uses thereof
WO2023086803A1 (en) 2021-11-10 2023-05-19 Iovance Biotherapeutics, Inc. Methods of expansion treatment utilizing cd8 tumor infiltrating lymphocytes
TW202337494A (en) 2021-11-16 2023-10-01 美商建南德克公司 Methods and compositions for treating systemic lupus erythematosus (sle) with mosunetuzumab
CA3235206A1 (en) 2021-11-16 2023-05-25 Davide BASCO Novel molecules for therapy and diagnosis
WO2023091968A1 (en) 2021-11-17 2023-05-25 Disc Medicine, Inc. Methods for treating anemia of kidney disease
WO2023099578A1 (en) 2021-12-01 2023-06-08 INSERM (Institut National de la Santé et de la Recherche Médicale) Neutralizing anti-cd95l monoclonal antibodies
WO2023114829A1 (en) 2021-12-15 2023-06-22 Genentech, Inc. Stabilized il-18 polypeptides and uses thereof
WO2023131901A1 (en) 2022-01-07 2023-07-13 Johnson & Johnson Enterprise Innovation Inc. Materials and methods of il-1beta binding proteins
WO2023139107A1 (en) 2022-01-18 2023-07-27 argenx BV Galectin-10 antibodies
TW202340251A (en) 2022-01-19 2023-10-16 美商建南德克公司 Anti-notch2 antibodies and conjugates and methods of use
WO2023147399A1 (en) 2022-01-27 2023-08-03 The Rockefeller University Broadly neutralizing anti-sars-cov-2 antibodies targeting the n-terminal domain of the spike protein and methods of use thereof
WO2023147488A1 (en) 2022-01-28 2023-08-03 Iovance Biotherapeutics, Inc. Cytokine associated tumor infiltrating lymphocytes compositions and methods
WO2023147486A1 (en) 2022-01-28 2023-08-03 Iovance Biotherapeutics, Inc. Tumor infiltrating lymphocytes engineered to express payloads
WO2023152637A1 (en) 2022-02-08 2023-08-17 INSERM (Institut National de la Santé et de la Recherche Médicale) Neutralization of acyl-coa binding protein confers autophagy-dependent organ protection
EP4227321A1 (en) 2022-02-10 2023-08-16 AVA Lifescience GmbH Antibodies targeting the b-cell receptor of chronic lymphocytic leukemia (cll) for use in treatment of cll
WO2023152204A1 (en) 2022-02-10 2023-08-17 Ava Lifescience Gmbh Antibodies targeting the b-cell receptor of chronic lymphocytic leukemia (cll) for use in treatment of cll
WO2023156437A1 (en) 2022-02-16 2023-08-24 INSERM (Institut National de la Santé et de la Recherche Médicale) Anti-robo4 human monoclonal antibodies and uses thereof for the treatment of cancer
WO2023156587A1 (en) 2022-02-18 2023-08-24 INSERM (Institut National de la Santé et de la Recherche Médicale) Use of tcr-deficient car-tregs in combination with anti-tcr complex monoclonal antibodies for inducing durable tolerance
WO2023161412A1 (en) 2022-02-25 2023-08-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies for preventing the cleavage of cd95l by metalloproteases
WO2023172883A1 (en) 2022-03-07 2023-09-14 Alpine Immune Sciences, Inc. Immunomodulatory proteins of variant cd80 polypeptides, cell therapies thereof and related methods and uses
WO2023170207A1 (en) 2022-03-09 2023-09-14 Alderaan Biotechnology Anti-cd160 transmembrane isoform antibodies
WO2023178192A1 (en) 2022-03-15 2023-09-21 Compugen Ltd. Il-18bp antagonist antibodies and their use in monotherapy and combination therapy in the treatment of cancer
WO2023178357A1 (en) 2022-03-18 2023-09-21 Evolveimmune Therapeutics, Inc. Bispecific antibody fusion molecules and methods of use thereof
US20240103010A1 (en) 2022-03-18 2024-03-28 Compugen Ltd. Pvrl2 and/or pvrig as biomarkers for treatment
US20230302423A1 (en) 2022-03-28 2023-09-28 Massachusetts Institute Of Technology Rna scaffolded wireframe origami and methods thereof
WO2023187657A1 (en) 2022-03-30 2023-10-05 Novartis Ag Methods of treating disorders using anti-natriuretic peptide receptor 1 (npr1) antibodies
WO2023192802A1 (en) 2022-04-01 2023-10-05 The Board Of Trustees Of The University Of Illinois Notch1 and notch4 decoys and methods of use
WO2023191816A1 (en) 2022-04-01 2023-10-05 Genentech, Inc. Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies
WO2023196877A1 (en) 2022-04-06 2023-10-12 Iovance Biotherapeutics, Inc. Treatment of nsclc patients with tumor infiltrating lymphocyte therapies
WO2023201299A1 (en) 2022-04-13 2023-10-19 Genentech, Inc. Pharmaceutical compositions of therapeutic proteins and methods of use
WO2023201369A1 (en) 2022-04-15 2023-10-19 Iovance Biotherapeutics, Inc. Til expansion processes using specific cytokine combinations and/or akti treatment
WO2023209177A1 (en) 2022-04-29 2023-11-02 Astrazeneca Uk Limited Sars-cov-2 antibodies and methods of using the same
WO2023215737A1 (en) 2022-05-03 2023-11-09 Genentech, Inc. Anti-ly6e antibodies, immunoconjugates, and uses thereof
WO2023220608A1 (en) 2022-05-10 2023-11-16 Iovance Biotherapeutics, Inc. Treatment of cancer patients with tumor infiltrating lymphocyte therapies in combination with an il-15r agonist
WO2023217743A1 (en) 2022-05-10 2023-11-16 Imcheck Therapeutics Anti-btn3a antibodies for use in methods of treating gastro-intestinal inflammatory disorders
WO2023219613A1 (en) 2022-05-11 2023-11-16 Genentech, Inc. Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies
WO2023217933A1 (en) 2022-05-11 2023-11-16 F. Hoffmann-La Roche Ag Antibody that binds to vegf-a and il6 and methods of use
WO2023222557A1 (en) 2022-05-17 2023-11-23 Bayer Aktiengesellschaft Radiopharmaceutical complexes targeting prostate-specific membrane antigen and its combinations
EP4279092A1 (en) 2022-05-17 2023-11-22 Bayer AG Radiopharmaceutical complexes targeting prostate-specific membrane antigen
WO2023228082A1 (en) 2022-05-26 2023-11-30 Pfizer Inc. Anti-tnfr2 antibodies and methods of use thereof
WO2023233330A1 (en) 2022-05-31 2023-12-07 Pfizer Inc. Anti-bmp9 antibodies and methods of use thereof
WO2023235699A1 (en) 2022-05-31 2023-12-07 Jounce Therapeutics, Inc. Antibodies to lilrb4 and uses thereof
WO2023240058A2 (en) 2022-06-07 2023-12-14 Genentech, Inc. Prognostic and therapeutic methods for cancer
WO2023239803A1 (en) 2022-06-08 2023-12-14 Angiex, Inc. Anti-tm4sf1 antibody-drug conjugates comprising cleavable linkers and methods of using same
WO2023245048A1 (en) 2022-06-15 2023-12-21 Bioverativ Usa Inc. Anti-complement c1s antibody formulation
WO2023242361A1 (en) 2022-06-15 2023-12-21 argenx BV Fcrn binding molecules and methods of use
WO2023250507A1 (en) 2022-06-24 2023-12-28 Bioverativ Usa Inc. Methods for treating complement-mediated diseases
WO2024011114A1 (en) 2022-07-06 2024-01-11 Iovance Biotherapeutics, Inc. Devices and processes for automated production of tumor infiltrating lymphocytes
WO2024015897A1 (en) 2022-07-13 2024-01-18 Genentech, Inc. Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies
WO2024020432A1 (en) 2022-07-19 2024-01-25 Genentech, Inc. Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies
WO2024020407A1 (en) 2022-07-19 2024-01-25 Staidson Biopharma Inc. Antibodies specifically recognizing b- and t-lymphocyte attenuator (btla) and uses thereof
EP4309733A1 (en) 2022-07-22 2024-01-24 Institut National de la Santé et de la Recherche Médicale (INSERM) Neutralization of acyl-coa binding protein for the treatment of cardiac dysfunction
WO2024020579A1 (en) 2022-07-22 2024-01-25 Bristol-Myers Squibb Company Antibodies binding to human pad4 and uses thereof
WO2024020564A1 (en) 2022-07-22 2024-01-25 Genentech, Inc. Anti-steap1 antigen-binding molecules and uses thereof
WO2024026496A1 (en) 2022-07-28 2024-02-01 Compugen Ltd. Combination therapy with anti-pvrig antibodies formulations and anti-pd-1 antibodies
WO2024030758A1 (en) 2022-08-01 2024-02-08 Iovance Biotherapeutics, Inc. Chimeric costimulatory receptors, chemokine receptors, and the use of same in cellular immunotherapies
WO2024030956A2 (en) 2022-08-03 2024-02-08 Mozart Therapeutics, Inc. Cd39-specific binding agents and methods of using the same
US20240052047A1 (en) 2022-08-03 2024-02-15 Pfizer Inc. Anti- il27r antibodies and methods of use thereof
WO2024028732A1 (en) 2022-08-05 2024-02-08 Janssen Biotech, Inc. Cd98 binding constructs for treating brain tumors
WO2024028731A1 (en) 2022-08-05 2024-02-08 Janssen Biotech, Inc. Transferrin receptor binding proteins for treating brain tumors
WO2024037633A2 (en) 2022-08-19 2024-02-22 Evive Biotechnology (Shanghai) Ltd Formulations comprising g-csf and uses thereof
WO2024044675A1 (en) 2022-08-25 2024-02-29 Beigene, Ltd. Methods of cancer treatment using anti-pd1 antibodies in combination with anti-tim3 antibodies
WO2024049949A1 (en) 2022-09-01 2024-03-07 Genentech, Inc. Therapeutic and diagnostic methods for bladder cancer
WO2024050524A1 (en) 2022-09-01 2024-03-07 University Of Georgia Research Foundation, Inc. Compositions and methods for directing apolipoprotein l1 to induce mammalian cell death
WO2024054929A1 (en) 2022-09-07 2024-03-14 Dynamicure Biotechnology Llc Anti-vista constructs and uses thereof
WO2024052503A1 (en) 2022-09-08 2024-03-14 Institut National de la Santé et de la Recherche Médicale Antibodies having specificity to ltbp2 and uses thereof
WO2024064752A2 (en) 2022-09-20 2024-03-28 Yale University Compositions of wet adhesives derived from vibrio cholerae biofilm adhesins and methods thereof
US20240101691A1 (en) 2022-09-21 2024-03-28 Sanofi Biotechnology Humanized anti-il-1r3 antibody and methods of use
WO2024074498A1 (en) 2022-10-04 2024-04-11 Imcheck Therapeutics Combination of a btn3a activating antibody, a bcl2 inhibitor and hypomethylating agent for use in treating cancer
WO2024077018A2 (en) 2022-10-04 2024-04-11 Alpine Immune Sciences, Inc. Methods and uses of taci-fc fusion immunomodulatory protein
WO2024077239A1 (en) 2022-10-07 2024-04-11 Genentech, Inc. Methods of treating cancer with anti-c-c motif chemokine receptor 8 (ccr8) antibodies

Citations (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US98193A (en) * 1869-12-21 Improvement in corn-planters
US158389A (en) * 1875-01-05 Improvement in gas-regulators
US5419904A (en) * 1990-11-05 1995-05-30 The Regents Of The University Of California Human B-lymphoblastoid cell line secreting anti-ganglioside antibody
US5576184A (en) * 1988-09-06 1996-11-19 Xoma Corporation Production of chimeric mouse-human antibodies with specificity to human tumor antigens
US5731168A (en) * 1995-03-01 1998-03-24 Genentech, Inc. Method for making heteromultimeric polypeptides
US5985599A (en) * 1986-05-29 1999-11-16 The Austin Research Institute FC receptor for immunoglobulin
US6165745A (en) * 1992-04-24 2000-12-26 Board Of Regents, The University Of Texas System Recombinant production of immunoglobulin-like domains in prokaryotic cells
US6277375B1 (en) * 1997-03-03 2001-08-21 Board Of Regents, The University Of Texas System Immunoglobulin-like domains with increased half-lives
US20030036459A1 (en) * 2001-08-16 2003-02-20 Jeon Ig-Kyun Brake system of vehicle
US6528624B1 (en) * 1998-04-02 2003-03-04 Genentech, Inc. Polypeptide variants
US6538124B1 (en) * 1998-04-02 2003-03-25 Genentech, Inc. Polypeptide variants
US20040002587A1 (en) * 2002-02-20 2004-01-01 Watkins Jeffry D. Fc region variants
US6676927B1 (en) * 1999-01-20 2004-01-13 The Rockefeller University Animal model and methods for its use in the selection of cytotoxic antibodies
US6737056B1 (en) * 1999-01-15 2004-05-18 Genentech, Inc. Polypeptide variants with altered effector function

Family Cites Families (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4752601A (en) 1983-08-12 1988-06-21 Immunetech Pharmaceuticals Method of blocking immune complex binding to immunoglobulin FC receptors
AU600575B2 (en) 1987-03-18 1990-08-16 Sb2, Inc. Altered antibodies
GB8916400D0 (en) 1989-07-18 1989-09-06 Dynal As Modified igg3
US5364930A (en) 1990-10-16 1994-11-15 Northwestern University Synthetic C1q peptide fragments
US5736137A (en) 1992-11-13 1998-04-07 Idec Pharmaceuticals Corporation Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma
EP0714409A1 (en) 1993-06-16 1996-06-05 Celltech Therapeutics Limited Antibodies
US6750334B1 (en) 1996-02-02 2004-06-15 Repligen Corporation CTLA4-immunoglobulin fusion proteins having modified effector functions and uses therefor
US5834597A (en) 1996-05-20 1998-11-10 Protein Design Labs, Inc. Mutated nonactivating IgG2 domains and anti CD3 antibodies incorporating the same
WO1998023289A1 (en) 1996-11-27 1998-06-04 The General Hospital Corporation MODULATION OF IgG BINDING TO FcRn
DE19721700C1 (en) 1997-05-23 1998-11-19 Deutsches Krebsforsch Mutant OKT3 antibody
PL199659B1 (en) 1998-02-25 2008-10-31 Merck Patent Gmbh Antibody-based fusion protein featured by prolonged residence in a cireculatory system and method of prolonging its residence time in a circulatory system
IL138608A0 (en) 1998-04-02 2001-10-31 Genentech Inc Antibody variants and fragments thereof
GB9809951D0 (en) 1998-05-08 1998-07-08 Univ Cambridge Tech Binding molecules
CA2341029A1 (en) 1998-08-17 2000-02-24 Abgenix, Inc. Generation of modified molecules with increased serum half-lives

Patent Citations (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US158389A (en) * 1875-01-05 Improvement in gas-regulators
US98193A (en) * 1869-12-21 Improvement in corn-planters
US5985599A (en) * 1986-05-29 1999-11-16 The Austin Research Institute FC receptor for immunoglobulin
US5576184A (en) * 1988-09-06 1996-11-19 Xoma Corporation Production of chimeric mouse-human antibodies with specificity to human tumor antigens
US5419904A (en) * 1990-11-05 1995-05-30 The Regents Of The University Of California Human B-lymphoblastoid cell line secreting anti-ganglioside antibody
US6165745A (en) * 1992-04-24 2000-12-26 Board Of Regents, The University Of Texas System Recombinant production of immunoglobulin-like domains in prokaryotic cells
US5731168A (en) * 1995-03-01 1998-03-24 Genentech, Inc. Method for making heteromultimeric polypeptides
US6277375B1 (en) * 1997-03-03 2001-08-21 Board Of Regents, The University Of Texas System Immunoglobulin-like domains with increased half-lives
US6821505B2 (en) * 1997-03-03 2004-11-23 Board Of Regents, The University Of Texas System Immunoglobin-like domains with increased half lives
US6528624B1 (en) * 1998-04-02 2003-03-04 Genentech, Inc. Polypeptide variants
US6538124B1 (en) * 1998-04-02 2003-03-25 Genentech, Inc. Polypeptide variants
US20040191244A1 (en) * 1999-01-15 2004-09-30 Genentech, Inc. Polypeptide variants with altered effector function
US20040228856A1 (en) * 1999-01-15 2004-11-18 Genentech, Inc. Polypeptide variants with altered effector function
US6737056B1 (en) * 1999-01-15 2004-05-18 Genentech, Inc. Polypeptide variants with altered effector function
US6676927B1 (en) * 1999-01-20 2004-01-13 The Rockefeller University Animal model and methods for its use in the selection of cytotoxic antibodies
US20030036459A1 (en) * 2001-08-16 2003-02-20 Jeon Ig-Kyun Brake system of vehicle
US20040002587A1 (en) * 2002-02-20 2004-01-01 Watkins Jeffry D. Fc region variants

Cited By (161)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030158389A1 (en) * 1998-04-02 2003-08-21 Genentech, Inc. Polypeptide variants
US7364731B2 (en) 1998-04-02 2008-04-29 Genentech, Inc. Polypeptide variants
US7297775B2 (en) 1998-04-02 2007-11-20 Genentech, Inc. Polypeptide variants
US7741072B2 (en) 1998-04-02 2010-06-22 Genentech, Inc. Polypeptide variants
US20060194954A1 (en) * 1998-04-02 2006-08-31 Genentech, Inc. Polypeptide variants
US20080274506A1 (en) * 1999-01-15 2008-11-06 Genentech, Inc. Polypeptide variants with altered effector function
US7335742B2 (en) 1999-01-15 2008-02-26 Genentech, Inc. Polypeptide variants with altered effector function
US8674083B2 (en) 1999-01-15 2014-03-18 Genentech, Inc. Polypeptide variants with altered effector function
US7332581B2 (en) 1999-01-15 2008-02-19 Genentech, Inc. Polypeptide variants with altered effector function
US8163882B2 (en) 1999-01-15 2012-04-24 Genentech, Inc. Polypeptide variants with altered effector function
US7371826B2 (en) 1999-01-15 2008-05-13 Genentech, Inc. Polypeptide variants with altered effector function
US20040191244A1 (en) * 1999-01-15 2004-09-30 Genentech, Inc. Polypeptide variants with altered effector function
US20050118174A1 (en) * 1999-01-15 2005-06-02 Genentech, Inc. Polypeptide variants with altered effector function
US7416727B2 (en) 1999-01-15 2008-08-26 Genentech, Inc. Polypeptide variants with altered effector function
US20060194957A1 (en) * 1999-01-15 2006-08-31 Genentech, Inc. Polypeptide variants with altered effector function
US20060194291A1 (en) * 1999-01-15 2006-08-31 Genentech, Inc. Polypeptide variants with altered effector function
US20060194290A1 (en) * 1999-01-15 2006-08-31 Genentech, Inc. Polypeptide variants with altered effector function
US20080274105A1 (en) * 1999-01-15 2008-11-06 Genentech, Inc. Polypeptide variants with altered effector function
US7122637B2 (en) 1999-01-15 2006-10-17 Genentech, Inc. Polypeptide variants with altered effector function
US7790858B2 (en) 1999-01-15 2010-09-07 Genentech, Inc. Polypeptide variants with altered effector function
US7785791B2 (en) 1999-01-15 2010-08-31 Genentech, Inc. Polypeptide variants with altered effector function
US20040228856A1 (en) * 1999-01-15 2004-11-18 Genentech, Inc. Polypeptide variants with altered effector function
US20080274108A1 (en) * 1999-01-15 2008-11-06 Genentech, Inc. Polypeptide variants with altered effector function
US7183387B1 (en) 1999-01-15 2007-02-27 Genentech, Inc. Polypeptide variants with altered effector function
US8545843B2 (en) 1999-05-07 2013-10-01 Genentech, Inc. Treatment of vasculitis
US9993550B2 (en) 1999-05-07 2018-06-12 Genentech, Inc. Treatment of pemphigus
US7820161B1 (en) 1999-05-07 2010-10-26 Biogen Idec, Inc. Treatment of autoimmune diseases
US8853366B2 (en) 2001-01-17 2014-10-07 Emergent Product Development Seattle, Llc Binding domain-immunoglobulin fusion proteins
US20080260731A1 (en) * 2002-03-01 2008-10-23 Bernett Matthew J Optimized antibodies that target cd19
US20070003546A1 (en) * 2002-03-01 2007-01-04 Xencor, Inc. Optimized Fc variants and methods for their generation
US8734791B2 (en) 2002-03-01 2014-05-27 Xencor, Inc. Optimized fc variants and methods for their generation
US20090068175A1 (en) * 2002-03-01 2009-03-12 Xencor, Inc. Optimized FC Variants and Methods for Their Generation
US7662925B2 (en) 2002-03-01 2010-02-16 Xencor, Inc. Optimized Fc variants and methods for their generation
US20080254027A1 (en) * 2002-03-01 2008-10-16 Bernett Matthew J Optimized CD5 antibodies and methods of using the same
US20090142340A1 (en) * 2002-03-01 2009-06-04 Xencor, Inc. Optimized Fc Variants and Methods for Their Generation
US8093357B2 (en) 2002-03-01 2012-01-10 Xencor, Inc. Optimized Fc variants and methods for their generation
US20080181890A1 (en) * 2002-03-01 2008-07-31 Xencor, Inc. Optimized Fc Variants and Methods for Their Generation
US8124731B2 (en) 2002-03-01 2012-02-28 Xencor, Inc. Optimized Fc variants and methods for their generation
US7317091B2 (en) 2002-03-01 2008-01-08 Xencor, Inc. Optimized Fc variants
US20070224189A1 (en) * 2002-03-01 2007-09-27 Xencor, Inc. CD20 OPTIMIZED Fc VARIANTS AND METHODS FOR THEIR GENERATION
US20070219133A1 (en) * 2002-03-01 2007-09-20 Xencor, Inc. CD52 OPTIMIZED Fc VARIANTS AND METHODS FOR THEIR GENERATION
US8093359B2 (en) 2002-09-27 2012-01-10 Xencor, Inc. Optimized Fc variants and methods for their generation
US20090092599A1 (en) * 2002-09-27 2009-04-09 Xencor, Inc. Optimized Fc variants and methods for their generation
US8753629B2 (en) 2002-09-27 2014-06-17 Xencor, Inc. Optimized Fc variants
US8735547B2 (en) 2002-09-27 2014-05-27 Xencor, Inc. Optimized Fc Variants
US8802823B2 (en) 2002-09-27 2014-08-12 Xencor, Inc. Optimized Fc variants
US8809503B2 (en) 2002-09-27 2014-08-19 Xencor, Inc. Optimized Fc variants and methods for their generation
US8858937B2 (en) 2002-09-27 2014-10-14 Xencor, Inc. Optimized Fc variants and methods for their generation
US8383109B2 (en) 2002-09-27 2013-02-26 Xencor, Inc. Optimized Fc variants and methods for their generation
US9193798B2 (en) 2002-09-27 2015-11-24 Xencor, Inc. Optimized Fc variants and methods for their generation
US9353187B2 (en) 2002-09-27 2016-05-31 Xencor, Inc. Optimized FC variants and methods for their generation
US8188231B2 (en) 2002-09-27 2012-05-29 Xencor, Inc. Optimized FC variants
US8039592B2 (en) 2002-09-27 2011-10-18 Xencor, Inc. Optimized Fc variants and methods for their generation
US20040132101A1 (en) * 2002-09-27 2004-07-08 Xencor Optimized Fc variants and methods for their generation
US20060235208A1 (en) * 2002-09-27 2006-10-19 Xencor, Inc. Fc variants with optimized properties
US10184000B2 (en) 2002-09-27 2019-01-22 Xencor, Inc. Optimized Fc variants and methods for their generation
US20070148171A1 (en) * 2002-09-27 2007-06-28 Xencor, Inc. Optimized anti-CD30 antibodies
US10183999B2 (en) 2002-09-27 2019-01-22 Xencor, Inc. Optimized Fc variants and methods for their generation
US8753628B2 (en) 2002-09-27 2014-06-17 Xencor, Inc. Optimized Fc variants
US20090081208A1 (en) * 2002-09-27 2009-03-26 Xencor, Inc. Optimized Fc variants and methods for their generation
US20070238665A1 (en) * 2003-03-03 2007-10-11 Xencor, Inc. Fc Variants Having Decreased Affinity for FcyRIIc
US9663582B2 (en) 2003-03-03 2017-05-30 Xencor, Inc. Optimized Fc variants
US8735545B2 (en) 2003-03-03 2014-05-27 Xencor, Inc. Fc variants having increased affinity for fcyrllc
US20070275460A1 (en) * 2003-03-03 2007-11-29 Xencor.Inc. Fc Variants With Optimized Fc Receptor Binding Properties
US10584176B2 (en) 2003-03-03 2020-03-10 Xencor, Inc. Fc variants with increased affinity for FcγRIIc
US20070248602A1 (en) * 2003-03-03 2007-10-25 Xencor, Inc. Fc Variants Having Increased Affinity for FcyRllc
US20070237766A1 (en) * 2003-03-03 2007-10-11 Xencor, Inc. Fc Variants Having Increased Affinity for FcyRllla
US8388955B2 (en) 2003-03-03 2013-03-05 Xencor, Inc. Fc variants
US20070248603A1 (en) * 2003-03-03 2007-10-25 Xencor, Inc. Fc Variants with Increased Affinity for FcyRlla
US10113001B2 (en) 2003-03-03 2018-10-30 Xencor, Inc. Fc variants with increased affinity for FcyRIIc
US20070237767A1 (en) * 2003-03-03 2007-10-11 Xencor, Inc. Fc Variants Having Decreased Affinity for FcyRllla
US9657106B2 (en) 2003-03-03 2017-05-23 Xencor, Inc. Optimized Fc variants
US20080057056A1 (en) * 2003-03-03 2008-03-06 Xencor, Inc. Fc Variants with Increased Affinity for FcyRIIC
US20070237765A1 (en) * 2003-03-03 2007-10-11 Xencor, Inc. Fc Variants Having Increased Affinity for FcyRl
US20070243188A1 (en) * 2003-03-03 2007-10-18 Xencor, Inc. Fc Variants Having Decreased Affinity for FcyRlla
US20110021755A1 (en) * 2003-03-03 2011-01-27 Xencor, Inc. Optimized Fc Variants
US8084582B2 (en) 2003-03-03 2011-12-27 Xencor, Inc. Optimized anti-CD20 monoclonal antibodies having Fc variants
US9051373B2 (en) 2003-05-02 2015-06-09 Xencor, Inc. Optimized Fc variants
US20050069521A1 (en) * 2003-08-28 2005-03-31 Emd Lexigen Research Center Corp. Enhancing the circulating half-life of interleukin-2 proteins
US9714282B2 (en) 2003-09-26 2017-07-25 Xencor, Inc. Optimized Fc variants and methods for their generation
US20060275283A1 (en) * 2003-11-12 2006-12-07 Biogen Idec Ma Inc. Fcgamma receptor-binding polypeptide variants and methods related thereto
US20100203046A1 (en) * 2003-11-12 2010-08-12 Biogen Idec Ma Inc. Fc gamma receptor-binding polypeptide variants and methods related thereto
US20100093979A1 (en) * 2003-12-22 2010-04-15 Gregory Alan Lazar Fc Polypeptides With Novel Fc Ligand Binding Sites
US20050249723A1 (en) * 2003-12-22 2005-11-10 Xencor, Inc. Fc polypeptides with novel Fc ligand binding sites
US8338575B2 (en) 2003-12-30 2012-12-25 Merck Patent Gmbh IL-7 fusion proteins
US7323549B2 (en) 2003-12-30 2008-01-29 Emd Lexigen Research Center Corp. IL-7 fusion proteins
US20050164352A1 (en) * 2003-12-30 2005-07-28 Emd Lexigen Research Center Corp. IL-7 fusion proteins
US20090010875A1 (en) * 2003-12-30 2009-01-08 Scott Lauder IL-7 Fusion Proteins
US7960514B2 (en) 2003-12-30 2011-06-14 Merck Patent Gmbh IL-7 fusion proteins
US7432357B2 (en) 2004-01-22 2008-10-07 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
JP2007537719A (en) * 2004-01-22 2007-12-27 メルク　パテント　ゲゼルシャフト　ミット　ベシュレンクテル　ハフトング Anticancer antibodies with reduced complement binding
US10633452B2 (en) 2004-01-22 2020-04-28 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US10017579B2 (en) 2004-01-22 2018-07-10 Meck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US9617349B2 (en) 2004-01-22 2017-04-11 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US20090148441A1 (en) * 2004-01-22 2009-06-11 Merck Patent Gmbh Anti-Cancer Antibodies With Reduced Complement Fixation
WO2005070967A2 (en) * 2004-01-22 2005-08-04 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
WO2005070967A3 (en) * 2004-01-22 2005-12-01 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US8835606B2 (en) 2004-01-22 2014-09-16 Merck Patent Gmbh Anti-cancer antibodies with reduced complement fixation
US20050202021A1 (en) * 2004-01-22 2005-09-15 Emd Lexigen Research Center Corp. Anti-cancer antibodies with reduced complement fixation
US20050244403A1 (en) * 2004-03-24 2005-11-03 Xencor, Inc. Immunoglobulin variants outside the Fc region
US7276585B2 (en) 2004-03-24 2007-10-02 Xencor, Inc. Immunoglobulin variants outside the Fc region
US20110064727A9 (en) * 2004-03-24 2011-03-17 Xencor, Inc. Immunoglobulin Variants Outside the Fc Region
US20080248028A1 (en) * 2004-03-24 2008-10-09 Xencor, Inc. Immunoglobulin Variants Outside the Fc Region
US11820830B2 (en) 2004-07-20 2023-11-21 Xencor, Inc. Optimized Fc variants
US20060074225A1 (en) * 2004-09-14 2006-04-06 Xencor, Inc. Monomeric immunoglobulin Fc domains
US8101720B2 (en) 2004-10-21 2012-01-24 Xencor, Inc. Immunoglobulin insertions, deletions and substitutions
US20060173170A1 (en) * 2004-11-12 2006-08-03 Xencor, Inc. Fc variants with altered binding to FcRn
US8367805B2 (en) 2004-11-12 2013-02-05 Xencor, Inc. Fc variants with altered binding to FcRn
US8802820B2 (en) 2004-11-12 2014-08-12 Xencor, Inc. Fc variants with altered binding to FcRn
US8318907B2 (en) 2004-11-12 2012-11-27 Xencor, Inc. Fc variants with altered binding to FcRn
US8546543B2 (en) 2004-11-12 2013-10-01 Xencor, Inc. Fc variants that extend antibody half-life
US11198739B2 (en) 2004-11-12 2021-12-14 Xencor, Inc. Fc variants with altered binding to FcRn
US8852586B2 (en) 2004-11-12 2014-10-07 Xencor, Inc. Fc variants with altered binding to FcRn
US8324351B2 (en) 2004-11-12 2012-12-04 Xencor, Inc. Fc variants with altered binding to FcRn
US8883973B2 (en) 2004-11-12 2014-11-11 Xencor, Inc. Fc variants with altered binding to FcRn
US9803023B2 (en) 2004-11-12 2017-10-31 Xencor, Inc. Fc variants with altered binding to FcRn
US10336818B2 (en) 2004-11-12 2019-07-02 Xencor, Inc. Fc variants with altered binding to FcRn
US8338574B2 (en) 2004-11-12 2012-12-25 Xencor, Inc. FC variants with altered binding to FCRN
US9200079B2 (en) 2004-11-12 2015-12-01 Xencor, Inc. Fc variants with altered binding to FcRn
US20060275282A1 (en) * 2005-01-12 2006-12-07 Xencor, Inc. Antibodies and Fc fusion proteins with altered immunogenicity
US10143748B2 (en) 2005-07-25 2018-12-04 Aptevo Research And Development Llc B-cell reduction using CD37-specific and CD20-specific binding molecules
US10307481B2 (en) 2005-07-25 2019-06-04 Aptevo Research And Development Llc CD37 immunotherapeutics and uses thereof
US9040041B2 (en) 2005-10-03 2015-05-26 Xencor, Inc. Modified FC molecules
US20100249382A1 (en) * 2005-10-03 2010-09-30 Xencor, Inc. MODIFIED Fc MOLECULES
US20080206867A1 (en) * 2005-10-03 2008-08-28 Desjarlais John R Fc variants with optimized Fc receptor binding properties
US7973136B2 (en) 2005-10-06 2011-07-05 Xencor, Inc. Optimized anti-CD30 antibodies
US20080267976A1 (en) * 2005-10-06 2008-10-30 Gregory Alan Lazar Optimized Anti-Cd30 Antibodies
US9574006B2 (en) 2005-10-06 2017-02-21 Xencor, Inc. Optimized anti-CD30 antibodies
US8409577B2 (en) 2006-06-12 2013-04-02 Emergent Product Development Seattle, Llc Single chain multivalent binding proteins with effector function
US9803020B2 (en) 2006-08-14 2017-10-31 Xencor, Inc. Optimized antibodies that target CD19
US8524867B2 (en) 2006-08-14 2013-09-03 Xencor, Inc. Optimized antibodies that target CD19
US10626182B2 (en) 2006-08-14 2020-04-21 Xencor, Inc. Optimized antibodies that target CD19
US20100272723A1 (en) * 2006-08-14 2010-10-28 Xencor, Inc. Optimized Antibodies that Target CD19
US11618788B2 (en) 2006-08-14 2023-04-04 Xencor, Inc. Optimized antibodies that target CD19
US20100104557A1 (en) * 2006-09-18 2010-04-29 Xencor, Inc. Optimized Antibodies that Target HM1.24
US9040042B2 (en) 2006-09-18 2015-05-26 Xencor, Inc. Optimized antibodies that target HM1.24
US8394374B2 (en) 2006-09-18 2013-03-12 Xencor, Inc. Optimized antibodies that target HM1.24
US8629245B2 (en) 2007-05-01 2014-01-14 Research Development Foundation Immunoglobulin Fc libraries
US10725037B2 (en) 2007-05-01 2020-07-28 Research Development Foundation Immunoglobulin Fc libraries
US20090136936A1 (en) * 2007-05-01 2009-05-28 George Georgiou Immunoglobulin fc libraries
US20080313379A1 (en) * 2007-06-15 2008-12-18 United Memories, Inc. Multiple bus charge sharing
US11932685B2 (en) 2007-10-31 2024-03-19 Xencor, Inc. Fc variants with altered binding to FcRn
US9101609B2 (en) 2008-04-11 2015-08-11 Emergent Product Development Seattle, Llc CD37 immunotherapeutic and combination with bifunctional chemotherapeutic thereof
US9238878B2 (en) 2009-02-17 2016-01-19 Redwood Bioscience, Inc. Aldehyde-tagged protein-based drug carriers and methods of use
US9879249B2 (en) 2009-02-17 2018-01-30 Redwood Bioscience, Inc. Aldehyde-tagged protein-based drug carriers and methods of use
US20100330076A1 (en) * 2009-06-30 2010-12-30 George Georgiou Immunoglobulin fc polypeptides
US8679493B2 (en) 2009-06-30 2014-03-25 Research Development Foundation Immunoglobulin Fc polypeptides
US11401348B2 (en) 2009-09-02 2022-08-02 Xencor, Inc. Heterodimeric Fc variants
US9475881B2 (en) 2010-01-19 2016-10-25 Xencor, Inc. Antibody variants with enhanced complement activity
US10183998B2 (en) 2011-01-14 2019-01-22 Redwood Bioscience, Inc. Aldehyde-tagged immunoglobulin polypeptides and methods of use thereof
US9540438B2 (en) 2011-01-14 2017-01-10 Redwood Bioscience, Inc. Aldehyde-tagged immunoglobulin polypeptides and methods of use thereof
US8969526B2 (en) 2011-03-29 2015-03-03 Roche Glycart Ag Antibody Fc variants
US10654916B2 (en) 2011-04-21 2020-05-19 The Regents Of The University Of California, A California Corporation Compositions and methods for the treatment of neuromyelitis optica
US11390667B2 (en) 2011-04-21 2022-07-19 The Regents Of The University Of California Compositions and methods for the treatment of neuromyelitis optica
US10683345B2 (en) 2012-07-13 2020-06-16 Roche Glycart Ag Bispecific anti-VEGF/anti-ANG-2 antibodies and their use in the treatment of ocular vascular diseases
US9695233B2 (en) 2012-07-13 2017-07-04 Roche Glycart Ag Bispecific anti-VEGF/anti-ANG-2 antibodies and their use in the treatment of ocular vascular diseases
US11332538B2 (en) 2015-02-09 2022-05-17 Research Development Foundation Engineered immunoglobulin Fc polypeptides displaying improved complement activation
US10457737B2 (en) 2015-02-09 2019-10-29 Research Development Foundation Engineered immunoglobulin Fc polypeptides displaying improved complement activation
US11352426B2 (en) 2015-09-21 2022-06-07 Aptevo Research And Development Llc CD3 binding polypeptides
US11208632B2 (en) 2016-04-26 2021-12-28 R.P. Scherer Technologies, Llc Antibody conjugates and methods of making and using the same
US11788066B2 (en) 2016-04-26 2023-10-17 R.P. Scherer Technologies, Llc Antibody conjugates and methods of making and using the same

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