US20080280322A9 - Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity - Google Patents
Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity Download PDFInfo
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- US20080280322A9 US20080280322A9 US11/199,232 US19923205A US2008280322A9 US 20080280322 A9 US20080280322 A9 US 20080280322A9 US 19923205 A US19923205 A US 19923205A US 2008280322 A9 US2008280322 A9 US 2008280322A9
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- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
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Definitions
- the present invention relates to the field of glycosylation engineering of proteins. More particularly, the present invention relates to glycosylation engineering to generate proteins with improved therapeutic properties, including antibodies with increased antibody-dependent cellular cytotoxicity.
- Glycoproteins mediate many essential functions in human beings, other eukaryotic organisms, and some prokaryotes, including catalysis, signaling, cell-cell communication, and molecular recognition and association. They make up the majority of non-cytosolic proteins in eukaryotic organisms. (Lis et al., Eur. J. Biochem. 218:1-27 (1993)). Many glycoproteins have been exploited for therapeutic purposes, and during the last two decades, recombinant versions of naturally-occurring, secreted glycoproteins have been a major product of the biotechnology industry.
- EPO erythropoietin
- therapeutic mAbs therapeutic monoclonal antibodies
- tPA tissue plasminogen activator
- IFN- ⁇ interferon- ⁇
- GM-CSF granulocyte-macrophage colony stimulating factor
- hCG human chorionic gonadotrophin
- the oligosaccharide component can significantly affect properties relevant to the efficacy of a therapeutic glycoprotein, including physical stability, resistance to protease attack, interactions with the immune system, pharmacokinetics, and specific biological activity. Such properties may depend not only on the presence or absence, but also on the specific structures, of oligosaccharides. Some generalizations between oligosaccharide structure and glycoprotein function can be made. For example, certain oligosaccharide structures mediate rapid clearance of the glycoprotein from the bloodstream through interactions with specific carbohydrate binding proteins, while others can be bound by antibodies and trigger undesired immune reactions. (Jenkins et al., Nature Biotechnol. 14:975-81 (1996)).
- Mammalian cells are the preferred hosts for production of therapeutic glycoproteins, due to their capability to glycosylate proteins in the most compatible form for human application. (Cumming et al., Glycobiology 1: 115-30 (1991); Jenkins et al., Nature Biotechnol. 14:975-81 (1996)). Bacteria very rarely glycosylate proteins, and like other types of common hosts, such as yeasts, filamentous fungi, insect and plant cells, yield glycosylation patterns associated with rapid clearance from the bloodstream, undesirable immune interactions, and in some specific cases, reduced biological activity. Among mammalian cells, Chinese hamster ovary (CHO) cells have been most commonly used during the last two decades.
- these cells allow consistent generation of genetically stable, highly productive clonal cell lines. They can be cultured to high densities in simple bioreactors using serum-free media, and permit the development of safe and reproducible bioprocesses.
- Other commonly used animal cells include baby hamster kidney (BHK) cells, NS0- and SP2/0-mouse myeloma cells. More recently, production from transgenic animals has also been tested (Jenkins et al., Nature Biotechnol. 14:975-81 (1996)).
- All antibodies contain carbohydrate structures at conserved positions in the heavy chain constant regions, with each isotype possessing a distinct array of N-linked carbohydrate structures, which variably affect protein assembly, secretion or functional activity.
- the structure of the attached N-linked carbohydrate varies considerably, depending on the degree of processing, and can include high-mannose, multiply-branched as well as biantennary complex oligosaccharides. (Wright, A., and Morrison, S. L., Trends Biotech. 15:26-32 (1997)).
- Unconjugated monoclonal antibodies can be useful medicines for the treatment of cancer, as demonstrated by the U.S. Food and Drug Administration's approval of Rituximab (RituxanTM; IDEC Pharmaceuticals, San Diego, Calif., and Genentech Inc., San Francisco, Calif.), for the treatment of CD20 positive B-cell, low-grade or follicular Non-Hodgkin's lymphoma, and Trastuzumab (HerceptinTM; Genentech Inc,) for the treatment of advanced breast cancer (Grillo-Lopez, A.-J., et al., Semin. Oncol. 26:66-73 (1999); Goldenberg, M. M., Clin. Ther.
- IgG1 type antibodies the most commonly used antibodies in cancer immunotherapy, are glycoproteins that have a conserved N-linked glycosylation site at Asn297 in each CH2 domain.
- ADCC antibody dependent cellular cytotoxicity
- the present inventors showed previously that overexpression in Chinese hamster ovary (CHO) cells of ⁇ (1,4)-N-acetylglucosaminyltransferase III (GnTIII), a glycosyltransferase catalyzing the formation of bisected oligosaccharides, significantly increases the in vitro ADCC activity of an anti-neuroblastoma chimeric monoclonal antibody (chCE7) produced by the engineered CHO cells.
- GnTIII Chinese hamster ovary
- the antibody chCE7 belongs to a large class of unconjugated mAbs which have high tumor affinity and specificity, but have too little potency to be clinically useful when produced in standard industrial cell lines lacking the GnTIII enzyme (Umana, P., et al., Nature Biotechnol. 17:176-180 (1999)). That study was the first to show that large increases of maximal in vitro ADCC activity could be obtained by increasing the proportion of constant region (Fc)-associated, bisected oligosaccharides above the levels found in naturally occurring antibodies.
- Fc constant region
- the present inventors have applied this technology to Rituximab, the anti-CD20, IDEC-C2B8 chimeric antibody.
- the present inventors have likewise applied the technology to the unconjugated anti-cancer mAb chG250
- the present inventors have now generated new glycosylation variants of the anti-CD20 monoclonal antibody (mAb) IDEC-C2B8 (Rituximab) and the anti-cancer mAb chG250 using genetically engineered mAb-producing cell lines that overexpress N-acetylglucosaminyltransferase III (GnTIII; EC 2.1.4.144) in a tetracycline regulated fashion.
- GnTIII is required for the synthesis of bisected oligosaccharides, which are found at low to intermediate levels in naturally-occurring human antibodies but are missing in mAbs produced in standard industrial cell lines.
- MabtheraTM the version of Rixtuximab marketed in Europe
- ADCC mouse-myeloma derived chG250 in biological activity.
- a ten-fold lower amount of the variant carrying the highest levels of bisected oligosaccharides was required to reach the maximal ADCC activity as MabtheraTM.
- the variant carrying the highest levels of bisected oligosaccharides mediated significant ADCC activity at a 125-fold lower concentration than that required to detect even low ADCC activity by the unmodified control chG250.
- a clear correlation was found between the level of GnTIII expression and ADCC activity.
- the claimed invention is directed to a host cell engineered to produce a polypeptide having increased Fc-mediated cellular cytotoxicity by expression of at least one nucleic acid encoding ⁇ (1,4)-N-acetylglucosaminyltransferase III (GnT III), wherein the polypeptide produced by the host cell is selected from the group consisting of a whole antibody molecule, an antibody fragment, and a fusion protein which includes a region equivalent to the Fc region of an immunoglobulin, and wherein the GnT III is expressed in an amount sufficient to increase the proportion of said polypeptide carrying bisected hybrid oligosaccharides or galactosylated complex oligosaccharides or mixtures thereof in the Fc region relative to polypeptides carrying bisected complex oligosaccharides in the Fc region.
- GnT III ⁇ (1,4)-N-acetylglucosaminyltransferase III
- the polypeptide is IgG or a fragment thereof, most preferably, IgG1 or a fragment thereof.
- the polypeptide is a fusion protein that includes a region equivalent to the Fc region of a human IgG.
- a nucleic acid molecule comprising at least one gene encoding GnTIII has been introduced into the host cell.
- at least one gene encoding GnTIII has been introduced into the host cell chromosome.
- the host cell has been engineered such that an endogenous GnT III gene is activated, for example, by insertion of a DNA element which increases gene expression into the host chromosome.
- the endogenous GnTIII has been activated by insertion of a promoter, an enhancer, a transcription factor binding site, a transposon, or a retroviral element or combinations thereof into the host cell chromosome.
- the host cell has been selected to carry a mutation triggering expression of an endogenous GnTIII.
- the host cell is the CHO cell mutant lec 10.
- the at least one nucleic acid encoding a GnTIII is operably linked to a constitutive promoter element.
- the host cell is a CHO cell, a BHK cell, a NS0 cell, a SP2/0 cell, or a hybridoma cell, a Y0 myeloma cell, a P3X63 mouse myeloma cell, a PER cell or a PER.C6 cell and said polypeptide is an anti-CD20 antibody.
- the host cell is a SP2/0 cell and the polypeptide is the monoclonal antibody chG250.
- the claimed invention is directed to a host cell that further comprises at least one transfected nucleic acid encoding an antibody molecule, an antibody fragment, or a fusion protein that includes a region equivalent to the Fc region of an immunoglobulin.
- the host cell comprises at least one transfected nucleic acid encoding an anti-CD20 antibody, the chimeric anti-human neuroblastoma monoclonal antibody chCE7, the chimeric anti-human renal cell carcinoma monoclonal antibody chG250, the chimeric anti-human colon, lung, and breast carcinoma monoclonal antibody ING-1, the humanized anti-human 17-1A antigen monoclonal antibody 3622W94, the humanized anti-human colorectal tumor antibody A33, the anti-human melanoma antibody directed against GD3 ganglioside R24, or the chimeric anti-human squamous-cell carcinoma monoclonal antibody SF-25, an anti-human EGFR antibody, an anti-human
- the claimed invention is directed to a method for producing a polypeptide in a host cell comprising culturing any of the above-described the host cells under conditions which permit the production of said polypeptide having increased Fc-mediated cellular cytotoxicity.
- the method further comprises isolating said polypeptide having increased Fc-mediated cellular cytotoxicity.
- the host cell comprises at least one nucleic acid encoding a fusion protein comprising a region equivalent to a glycosylated Fc region of an immunoglobulin.
- the proportion of bisected oligosaccharides in the Fc region of said polypeptides is greater than 50%, more preferably, greater than 70%. In another embodiment, the proportion of bisected hybrid oligosaccharides or galactosylated complex oligosaccharides or mixtures thereof in the Fc region is greater than the proportion of bisected complex oligosaccharides in the Fc region of said polypeptide.
- the polypeptide is an anti-CD20 antibody and the anti-CD20 antibodies produced by said host cell have a glycosylation profile, as analyzed by MALDI/TOF-MS, that is substantially equivalent to that shown in FIG. 2E .
- the polypeptide is the chG250 monoclonal antibody and the chG250 antibodies produced by said host cell have a glycosylaton profile, as analyzed by MALDI/TOF-MS, that is substantially equivalent to that shown in FIG. 7D .
- the claimed invention is directed to an antibody having increased antibody dependent cellular cytotoxicity (ADCC) produced by any of the methods described above.
- ADCC antibody dependent cellular cytotoxicity
- the antibody is selected from the group consisting of an anti-CD20 antibody, chCE7, ch-G250, a humanized anti-HER2 monoclonal antibody, ING-1, 3622W94, SP-25, A33, and R24.
- the polypeptide can be an antibody fragment that includes a region equivalent to the Fc region of an immunoglobulin, having increased Fc-mediated cellular cytotoxicity produced by any of the methods described above.
- the claimed invention is directed to a fusion protein that includes a region equivalent to the Fc region of an immunoglobulin and having increased Fc-mediated cellular cytotoxicity produced by any of the methods described above.
- the claimed invention is directed to a pharmaceutical composition
- a pharmaceutical composition comprising the antibody, antibody fragment, or fusion protein of the invention and a pharmaceutically acceptable carrier.
- the claimed invention is directed to a method for the treatment of cancer comprising administering a therapeutically effective amount of said pharmaceutical composition to a patient in need thereof.
- the invention is directed to an improved method for treating an autoimmune disease produced in whole or in part by pathogenic autoantibodies based on B-cell depletion comprising administering a therapeutically effective amount of immunologically active antibody to a human subject in need thereof, the improvement comprising administering a therapeutically effective amount of an antibody having increased ADCC prepared as described above.
- the antibody is an anti-CD20 antibody.
- autoimmune diseases or disorders include, but are not limited to, immune-mediated thrombocytopenias, such as acute idiopathic thrombocytopenic purpurea and chronic idiopathic thrombocytopenic purpurea, dermatomyositis, Sydenham's chorea, lupus nephritis, rheumatic fever, polyglandular syndromes, Henoch-Schonlein purpura, post-streptococcal nephritis, erythema nodosum, Takayasu's arteritis, Addison's disease, erythema multiforme, polyarteritis nodosa, ankylosing spondylitis, Goodpasture's syndrome, thromboangitis ubiterans, primary biliary cirrhosis, Hashimoto's thyroiditis, thyrotoxicosis, chronic active hepatitis, polymyositis/
- atopic dermatitis atopic dermatitis
- systemic scleroderma and sclerosis responses associated with inflammatory bowel disease (such as Crohn's disease and ulcerative colitis); respiratory distress syndrome (including adult respiratory distress syndrome; ARDS); dermatitis; meningitis; encephalitis; uveitis; colitis; glomerulonephritis; allergic conditions such as eczema and asthma and other conditions involving infiltration of T cells and chronic inflammatory responses; atherosclerosis; leukocyte adhesion deficiency; rheumatoid arthritis; systemic lupus erythematosus (SLE); diabetes mellitus (e.g.
- FIG. 1 Indirect immunofluorescence assay showing the reactivity of the antibody preparation C2B8-25t to CD20 positive SB cells. Negative controls, including the HSB CD20 negative cell line and cells treated only with the secondary FITC-conjugated anti-human Fc polyclonal antibody are not shown.
- FIG. 2A-2E MALDI/TOF-MS spectra of the oligosaccharides derived from MabtheraTM ( FIG. 2A ), C2B8-nt ( FIG. 2B ), C2B8-2000t ( FIG. 2C ), C2B8-50t ( FIG. 2D ), and C2B8-25t ( FIG. 2E ) antibody samples. Oligosaccharides appear as [M+Na + ] and [M+K + ] ions.
- Oligosaccharide appearing in the first two spectra were derived from cell cultures that do not express GnTIII, whereas oligosaccharides in C, D, and E were derived from a single cell line expressing GnTIII at different levels (i.e. tetracycline concentrations).
- FIGS. 3A and 3B Illustration of a typical human IgG Fc-associated oligosaccharide structure (A) and partial N-linked glycosylation pathway (B).
- FIG. 3A The core of the oligosaccharide is composed of three mannose (M) and two N-acetylglucosamine (Gn) monosaccharide residues attached to Asn 297 .
- M mannose
- Gn N-acetylglucosamine
- G fucose
- N bisecting N-acetylglucosamine
- Terminal N-acetylneuraminic acid maybe also present but it is not included in the figure.
- FIGS. 4A and 4B ADCC activities of Rituximab glycosylation variants. The percentage of cytotoxicity was measured via lysis of 51 Cr labeled CD20-positive SB cells by human lymphocytes (E:T ratio of 100:1) mediated by different mAb concentrations.
- FIG. 4A Activity of C2B8 samples derived from a single cell line but produced at increasing GnTIII expression levels (i.e., decreasing tetracycline concentrations). The samples are C2B8-2000t, C2B8-50t, C2B8-25t, and C2B8-nt (control mAb derived from a clone that does not express GnTIII
- FIG. 4B ADCC activity of C2B8-50t and C2B8-25t compared to MabtheraTM.
- FIG. 5 Western blot analysis of the seven GnTIII expressing clones and the wild type. 30 ⁇ g of each sample were loaded on a 8.75% SDS gel, transferred to a PVDF membrane and probed with the anti-c-myc monoclonal antibody (9E10). WT refers to wt-chG250-SP2/0 cells.
- FIG. 6 SDS polyacrylamide gel electrophoresis of resolved purified antibody samples.
- FIG. 7A-7D MALDI/TOF-MS spectra of neutral oligosaccharide mixtures from chG250 mAb samples produced by clones expressing different GnTIII levels and wt-chG250-SP2/0 cells: WT ( FIG. 7A ), 2F1 ( FIG. 7B ), 3D3 ( FIG. 7C ), 4E6 ( FIG. 7D ).
- FIG. 8A-8D MALDI/TOF-MS spectra of neutral oligosaccharide mixtures from chG250 mAb samples produced by clones expressing different GnTIII levels: 4E8, ( FIG. 8A ); 5G2, ( FIG. 8B ); 4G3, ( FIG. 8C ); 5H12, ( FIG. 8D ).
- FIG. 9 In vitro ADCC assay of antibody samples derived from control wt-chG250-SP2/-cells and GnTIII transected clones 3D3 and 5H12.
- antibody is intended to include whole antibody molecules, antibody fragments, or fusionproteins that include a region equivalent to the Fc region of an immunoglobulin.
- region equivalent to the Fc region of an immunoglobulin is intended to include naturally occurring allelic variants of the Fc region of an immunoglobulin as well as variants having alterations which produce substitutions, additions, or deletions but which do not decrease substantially the ability of the immunoglobulin to mediate antibody dependent cellular cytotoxicity.
- one or more amino acids can be deleted from the N-terminus or C-terminus of the Fc region of an immunoglobulin without substantial loss of biological function.
- variants can be selected according to general rules known in the art so as to have minimal effect on activity. (See, e.g., Bowie, J. U. et al., Science 247:1306-10 (1990).
- glycoprotein-modifying glycosyl transferase refers to ⁇ (1,4)-N-acetylglucosaminyltransferase III (GnTIII).
- glycosylation engineering includes metabolic engineering of the glycosylation machinery of a cell, including genetic manipulations of the oligosaccharide synthesis pathways to achieve altered glycosylation of glycoproteins expressed in cells. Furthermore, glycosylation engineering includes the effects of mutations and cell environment on glycosylation.
- host cell covers any kind of cellular system which can be engineered to generate modified glycoforms of proteins, protein fragments, or peptides of interest, including antibodies and antibody fragments.
- the host cells have been manipulated to express optimized levels of GnT III.
- Host cells include cultured cells, e.g., mammalian cultured cells, such as CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells or hybridoma cells, yeast cells, and insect cells, to name only a few, but also cells comprised within a transgenic animal or cultured tissue.
- Fc-mediated cellular cytotoxicity includes antibody-dependent cellular cytotoxicity and cellular cytotoxicity mediated by a soluble Fc-fusion protein containing a human Fc-region. It is an immune mechanism leading to the lysis of “antibody-targeted cells” by “human immune effector cells”, wherein:
- the term increased Fc-mediated cellular cytotoxicity is defined as either an increase in the number of “antibody-targeted cells” that are lysed in a given time, at a given concentration of antibody, or of Fc-fusion protein, in the medium surrounding the target cells, by the mechanism of Fc-mediated cellular cytotoxicity defined above, and/or a reduction in the concentration of antibody, or of Fc-fusion protein, in the medium surrounding the target cells, required to achieve the lysis of a given number of “antibody-targeted cells”, in a given time, by the mechanism of Fc-mediated cellular cytotoxicity.
- Fc-mediated cellular cytotoxicity is relative to the cellular cytotoxicity mediated by the same antibody, or Fc-fusion protein, produced by the same type of host cells, using the same standard production, purification, formulation and storage methods, which are known to those skilled in the art, but that has not been produced by host cells engineered to express the glycosyltransferase GnTIII by the methods described herein.
- ADCC antibody dependent cellular cytotoxicity
- anti-CD20 antibody is intended to mean an antibody which specifically recognizes a cell surface non-glycosylated phosphoprotein of 35,000 Daltons, typically designated as the human B lymphocyte restricted differentiation antigen Bp35, commonly referred to as CD20.
- the present invention provides methods for the generation and use of host cell systems for the production of glycoforms of antibodies or antibody fragments or fusion proteins which include antibody fragments with increased antibody-dependent cellular cytotoxicity. Identification of target epitopes and generation of antibodies having potential therapeutic value, for which modification of the glycosylation pattern is desired, and isolation of their respective coding nucleic acid sequence is within the scope of the invention.
- antibodies to target epitopes of interest include but are not limited to polyclonal, monoclonal, chimeric, single chain, Fab fragments and fragments produced by an Fab expression library.
- Such antibodies may be useful, e.g., as diagnostic or therapeutic agents.
- therapeutic agents neutralizing antibodies, i.e., those which compete for binding with a ligand, substrate or adapter molecule, are of especially preferred interest.
- various host animals are immunized by injection with the target protein of interest including, but not limited to, rabbits, mice, rats, etc.
- Various adjuvants may be used to increase the immunological response, depending on the host species, including but not limited to Freund's (complete and incomplete), mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, saponin, oil emulsions, keyhole limpet hemocyanin, dinitrophenol, and potentially useful human adjuvants such as BCG (bacille Calmette-Guerin) and Corynebacterium parvum.
- BCG Bacille Calmette-Guerin
- Monoclonal antibodies to the target of interest may be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique originally described by Kohler and Milstein, Nature 256:495-97 (1975), the human B-cell hybridoma technique (Kosbor et al., Immunology Today 4:72 (1983); Cote et al., Proc. Natl. Acad. Sci. U.S.A. 80:2026-30 (1983) and the EBV-hybridoma technique (Cole et al., Monoclonal Antibodies and Cancer Therapy 77-96 (Alan R. Liss, Inc., 1985)).
- Antibody fragments which contain specific binding sites of the target protein of interest may be generated by known techniques.
- such fragments include, but are not limited to, F(ab′) 2 fragments which can be produced by pepsin digestion of the antibody molecule and the Fab fragments which can be generated by reducing the disulfide bridges of the F(ab′) 2 fragments.
- Fab expression libraries may be constructed (Huse et al., Science 246:1275-81 (1989) to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity to the target protein of interest.
- the coding nucleic acid sequence is identified and isolated using techniques well known in the art.
- the present invention provides host cell expression systems for the generation of proteins having modified glycosylation patterns.
- the present invention provides host cell systems for the generation of glycoforms of proteins having an improved therapeutic value. Therefore, the invention provides host cell expression systems selected or engineered to increase the expression of a glycoprotein-modifying glycosyltransferase, namely ⁇ (1,4)-N-acetylglucosaminyltransferase III (GnTIII).
- GnTIII glycoprotein-modifying glycosyltransferase
- such host cell expression systems may be engineered to comprise a recombinant nucleic acid molecule encoding GnTIII, operatively linked to a constitutive or regulated promoter system.
- host cell expression systems may be employed that naturally produce, are induced to produce, and/or are selected to produce GnTIII.
- the present invention provides a host cell that has been engineered to express at least one nucleic acid encoding GnTIII.
- the host cell is transformed or transfected with a nucleic acid molecule comprising at least one gene encoding GnTIII.
- the host cell has been engineered and/or selected in such way that endogenous GnTIII is activated.
- the host cell may be selected to carry a mutation triggering expression of endogenous GnTIII.
- the host cell is a CHO lec10 mutant.
- the host cell may be engineered such that endogenous GnTIII is activated.
- the host cell is engineered such that endogenous GnTIII has been activated by insertion of a constitutive promoter element, a transposon, or a retroviral element into the host cell chromosome.
- any type of cultured cell line can be used as a background to engineer the host cell lines of the present invention.
- CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells or hybridoma cells, yeast cells, or insect cells are used as the background cell line to generate the engineered host cells of the invention.
- the invention is contemplated to encompass any engineered host cells expressing GnTIII as defined herein.
- nucleic acids encoding GnTIII may be expressed under the control of a constitutive promoter or, alternately, a regulated expression system.
- Suitable regulated expression systems include, but are not limited to, a tetracycline-regulated expression system, an ecdysone-inducible expression system, a lac-switch expression system, a glucocorticoid-inducible expression system, a temperature-inducible promoter system, and a metallothionein metal-inducible expression system. If several different nucleic acids encoding GnTIII are comprised within the host cell system, some of them may be expressed under the control of a constitutive promoter, while others are expressed under the control of a regulated promoter.
- the maximal expression level is considered to be the highest possible level of stable GnTIII expression that does not have a significant adverse effect on cell growth rate, and will be determined using routine experimentation.
- Expression levels are determined by methods generally known in the art, including Western blot analysis using a GnTIII specific antibody, Northern blot analysis using a GnTIII specific nucleic acid probe, or measurement of enzymatic activity.
- a lectin may be employed which binds to biosynthetic products of the GnTIII, for example, E 4 -PHA lectin.
- the nucleic acid may be operatively linked to a reporter gene; the expression levels of the GnTIII are determined by measuring a signal correlated with the expression level of the reporter gene.
- the reporter gene may transcribed together with the nucleic acid(s) encoding said GnTIII as a single mRNA molecule; their respective coding sequences may be linked either by an internal ribosome entry site (IRES) or by a cap-independent translation enhancer (CITE).
- the reporter gene may be translated together with at least one nucleic acid encoding said GnTIII such that a single polypeptide chain is formed.
- the nucleic acid encoding the GnTIII may be operatively linked to the reporter gene under the control of a single promoter, such that the nucleic acid encoding the GnTIII and the reporter gene are transcribed into an RNA molecule which is alternatively spliced into two separate messenger RNA (mRNA) molecules; one of the resulting mRNAs is translated into said reporter protein, and the other is translated into said GnTIII.
- mRNA messenger RNA
- nucleic acids encoding GnTIII may be arranged in such way that they are transcribed as one or as several mRNA molecules. If they are transcribed as a single mRNA molecule, their respective coding sequences may be linked either by an internal ribosome entry site (IRES) or by a cap-independent translation enhancer (CITE). They may be transcribed from a single promoter into an RNA molecule which is alternatively spliced into several separate messenger RNA (mRNA) molecules, which then are each translated into their respective encoded GnTIII.
- IRS internal ribosome entry site
- CITE cap-independent translation enhancer
- the present invention provides host cell expression systems for the generation of therapeutic antibodies, having an increased antibody-dependent cellular cytotoxicity, and cells which display the IgG Fc region on the surface to promote Fc-mediated cytotoxicity.
- the host cell expression systems have been engineered and/or selected to express nucleic acids encoding the antibody for which the production of altered glycoforms is desired, along with at least one nucleic acid encoding GnTIII.
- the host cell system is transfected with at least one gene encoding GnTIII.
- the transfected cells are selected to identify and isolate clones that stably express the GnTIII.
- the host cell has been selected for expression of endogenous GnTIII.
- cells may be selected carrying mutations which trigger expression of otherwise silent GnTIII.
- CHO cells are known to carry a silent GnT III gene that is active in certain mutants, e.g., in the mutant Lec10.
- methods known in the art may be used to activate silent GnTIII, including the insertion of a regulated or constitutive promoter, the use of transposons, retroviral elements, etc.
- gene knockout technologies or the use of ribozyme methods may be used to tailor the host cell's GnTIII expression level, and is therefore within the scope of the invention.
- any type of cultured cell line can be used as background to engineer the host cell lines of the present invention.
- CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells or hybridoma cells, yeast cell, or insect cells may be used.
- such cell lines are engineered to further comprise at least one transfected nucleic acid encoding a whole antibody molecule, an antibody fragment, or a fusion protein that includes a region equivalent to the Fc region of an immunoglobulin.
- a hybridoma cell line expressing a particular antibody of interest is used as background cell line to generate the engineered host cells of the invention.
- At least one nucleic acid in the host cell system encodes GnT III.
- One or several nucleic acids encoding GnTIII may be expressed under the control of a constitutive promoter, or alternately, a regulated expression system.
- Suitable regulated expression systems include, but are not limited to, a tetracycline-regulated expression system, an ecdysone-inducible expression system, a lac-switch expression system, a glucocorticoid-inducible expression system, a temperature-inducible promoter system, and a metallothionein metal-inducible expression system. If several different nucleic acids encoding GnTIII are comprised within the host cell system, some of them may be expressed under the control of a constitutive promoter, while others are expressed under the control of a regulated promoter.
- the maximal expression level is considered to be the highest possible level of stable GnTIII expression that does not have a significant adverse effect on cell growth rate, and will be determined using routine experimentation.
- Expression levels are determined by methods generally known in the art, including Western blot analysis using a GnTIII specific antibody, Northern blot analysis using a GnTIII specific nucleic acid probe, or measurement of GnTIII enzymatic activity.
- a lectin may be employed which binds to biosynthetic products of GnTIII, for example, E 4 -PHA lectin.
- the nucleic acid may be operatively linked to a reporter gene; the expression levels of the glycoprotein-modifying glycosyl transferase are determined by measuring a signal correlated with the expression level of the reporter gene.
- the reporter gene may transcribed together with the nucleic acid(s) encoding said glycoprotein-modifying glycosyl transferase as a single mRNA molecule; their respective coding sequences may be linked either by an internal ribosome entry site (IRES) or by a cap-independent translation enhancer (CITE).
- the reporter gene may be translated together with at least one nucleic acid encoding GnTIII such that a single polypeptide chain is formed.
- the nucleic acid encoding the GnTIII may be operatively linked to the reporter gene under the control of a single promoter, such that the nucleic acid encoding the GnTIII and the reporter gene are transcribed into an RNA molecule which is alternatively spliced into two separate messenger RNA (mRNA) molecules; one of the resulting mRNAs is translated into said reporter protein, and the other is translated into said GnTIII.
- mRNA messenger RNA
- nucleic acids encoding a GnTIII may be arranged in such way that they are transcribed as one or as several mRNA molecules. If they are transcribed as single mRNA molecule, their respective coding sequences may be linked either by an internal ribosome entry site (IRES) or by a cap-independent translation enhancer (CITE). They may be transcribed from a single promoter into an RNA molecule which is alternatively spliced into several separate messenger RNA (mRNA) molecules, which then are each translated into their respective encoded GnTIII.
- IRS internal ribosome entry site
- CITE cap-independent translation enhancer
- Expression Systems which are well known to those skilled in the art can be used to construct expression vectors containing the coding sequence of the protein of interest and the coding sequence of the GnTIII and appropriate transcriptional/translational control signals. These methods include in vitro recombinant DNA techniques, synthetic techniques and in vivo recombination/genetic recombination. See, for example, the techniques described in Maniatis et al., Molecular Cloning A Laboratory Manual, Cold Spring Harbor Laboratory, N.Y. (1989) and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, N.Y (1989).
- host-expression vector systems may be utilized to express the coding sequence of the protein of interest and the coding sequence of the GnTIII.
- mammalian cells are used as host cell systems transfected with recombinant plasmid DNA or cosmid DNA expression vectors containing the coding sequence of the protein of interest and the coding sequence of the GnTIII.
- CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER. C6 cells or hybridoma cells, yeast cells, or insect cells are used as host cell system.
- yeast cells transformed with recombinant yeast expression vectors containing the coding sequence of the protein of interest and the coding sequence of the GnTIII include, yeast cells transformed with recombinant yeast expression vectors containing the coding sequence of the protein of interest and the coding sequence of the GnTIII; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing the coding sequence of the protein of interest and the coding sequence of the GnTIII; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing the coding sequence of the protein of interest and the coding sequence of the GnTIII; or animal cell systems infected with recombinant virus expression vectors (e.g., adenovirus, vaccinia virus) including cell lines engineered to contain multiple copies
- stable expression is generally preferred to transient expression because it typically achieves more reproducible results and also is more amenable to large scale production.
- host cells can be transformed with the respective coding nucleic acids controlled by appropriate expression control elements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.), and a selectable marker.
- appropriate expression control elements e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.
- engineered cells may be allowed to grow for 1-2 days in an enriched media, and then are switched to a selective media.
- the selectable marker in the recombinant plasmid confers resistance to the selection and allows selection of cells which have stably integrated the plasmid into their chromosomes and grow to form foci which in turn can be cloned and expanded into cell lines.
- a number of selection systems may be used, including, but not limited to, the herpes simplex virus thymidine kinase (Wigler et al., Cell 11:223 (1977)), hypoxanthine-guanine phosphoribosyltransferase (Szybalska & Szybalski, Proc. Natl. Acad. Sci. USA 48:2026 (1962)), and adenine phosphoribosyltransferase (Lowy et al., Cell 22:817 (1980)) genes, which can be employed in tk ⁇ , hgprt ⁇ or aprt ⁇ cells, respectively.
- antimetabolite resistance can be used as the basis of selection for dhfr, which confers resistance to methotrexate (Wigler et al., Natl. Acad. Sci. USA 77:3567 (1989); O'Hare et al., Proc. Natl. Acad. Sci. USA 78:1527 (1981)); gpt, which confers resistance to mycophenolic acid (Mulligan & Berg, Proc. Natl. Acad. Sci. USA 78:2072 (1981)); neo, which confers resistance to the aminoglycoside G-418 (Colberre-Garapin et al., J. Mol. Biol.
- trpB which allows cells to utilize indole in place of tryptophan
- hisD which allows cells to utilize histinol in place of histidine
- the host cells which contain the coding sequence and which express the biologically active gene products may be identified by at least four general approaches; (a) DNA-DNA or DNA-RNA hybridization; (b) the presence or absence of “marker” gene functions; (c) assessing the level of transcription as measured by the expression of the respective mRNA transcripts in the host cell; and (d) detection of the gene product as measured by immunoassay or by its biological activity.
- the presence of the coding sequence of the protein of interest and the coding sequence of the GnTIII inserted in the expression vector can be detected by DNA-DNA or DNA-RNA hybridization using probes comprising nucleotide sequences that are homologous to the respective coding sequences, respectively, or portions or derivatives thereof.
- the recombinant expression vector/host system can be identified and selected based upon the presence or absence of certain “marker” gene functions (e.g., thymidine kinase activity, resistance to antibiotics, resistance to methotrexate, transformation phenotype, occlusion body formation in baculovirus, etc.).
- certain “marker” gene functions e.g., thymidine kinase activity, resistance to antibiotics, resistance to methotrexate, transformation phenotype, occlusion body formation in baculovirus, etc.
- certain “marker” gene functions e.g., thymidine kinase activity, resistance to antibiotics, resistance to methotrexate, transformation phenotype, occlusion body formation in baculovirus, etc.
- a marker gene can be placed in tandem with the coding sequences under the control of the same or different promoter used to control the expression of the coding sequences. Expression of the marker in response to induction
- transcriptional activity for the coding region of the protein of interest and the coding sequence of the GnTIII can be assessed by hybridization assays.
- RNA can be isolated and analyzed by Northern blot using a probe homologous to the coding sequences of the protein of interest and the coding sequence of the GnTIII or particular portions thereof.
- total nucleic acids of the host cell may be extracted and assayed for hybridization to such probes.
- the expression of the protein products of the protein of interest and the coding sequence of the GnTIII can be assessed immunologically, for example by Western blots, immunoassays such as radioimmuno-precipitation, enzyme-linked immunoassays and the like.
- the ultimate test of the success of the expression system involves the detection of the biologically active gene products.
- the present invention provides glycoforms of antibodies and antibody fragments having increased antibody-dependent cellular cytotoxicity.
- the antigens of these two mAbs are highly expressed in their respective tumor cells and the antibodies mediate potent tumor destruction by effector cells in vitro and in vivo.
- many other unconjugated mAbs with fine tumor specificities cannot trigger effector functions of sufficient potency to be clinically useful.
- adjunct cytokine therapy is currently being tested. Addition of cytokines can stimulate antibody-dependent cellular cytotoxicity (ADCC) by increasing the activity and number of circulating lymphocytes.
- ADCC antibody-dependent cellular cytotoxicity
- ADCC a lytic attack on antibody-targeted cells, is triggered upon binding of leukocyte receptors to the constant region (Fc) of antibodies. Deo et al., Immunology Today 18:127 (1997).
- Fc ⁇ Rs lymphocyte receptors
- An IgG molecule carries two N-linked oligosaccharides in its Fc region, one on each heavy chain.
- an antibody is produced as a population of glycoforms which share the same polypeptide backbone but have different oligosaccharides attached to the glycosylation sites.
- the oligosaccharides normally found in the Fc region of serum IgG are of complex bi-antennary type (Wormald et al., Biochemistry 36:130-38 (1997), with low level of terminal sialic acid and bisecting N-acetylglucosamine (GlcNAc), and a variable degree of terminal galactosylation and core fucosylation.
- IgGs expressed in these cell lines lack, however, the bisecting GlcNAc found in low amounts in serum IgGs. Lifely et al., Glycobiology 318:813-22 (1995).
- CAMPATH-1H humanized IgG1
- the rat cell-derived antibody reached a similar in vitro ADCC activity as CAMPATH-1H antibodies produced in standard cell lines, but at significantly lower antibody concentrations.
- the CAMPATH antigen is normally present at high levels on lymphoma cells, and this chimeric mAb has high ADCC activity in the absence of a bisecting GlcNAc. Lifely et al., Glycobiology 318:813-22 (1995). In the N-linked glycosylation pathway, a bisecting GlcNAc is added by the enzyme ⁇ (1,4)-N-acetylglucosaminyltransferase III (GnT III). Schachter, Biochem. Cell Biol. 64:163-81 (1986).
- the present inventors used a single antibody-producing CHO cell line, that was previously engineered to express, in an externally-regulated fashion, different levels of a cloned GnT III gene. This approach established for the first time a rigorous correlation between expression of GnTIII and the ADCC activity of the modified antibody.
- C2B8 antibody modified according to the disclosed method had an about sixteen-fold higher ADCC activity than the standard, unmodified C2B8 antibody produced under identical cell culture and purification conditions.
- a C2B8 antibody sample expressed in CHO-tTA-C2B8 cells that do not have GnT III expression showed a cytotoxic activity of about 31% (at 1 ⁇ g/ml antibody concentration), measured as in vitro lysis of SB cells (CD20+) by human lymphocytes.
- C2B8 antibody derived from a CHO cell culture expressing GnT III at a basal, largely repressed level showed at 1 ⁇ g/ml antibody concentration a 33% increase in ADCC activity against the control at the same antibody concentration.
- antibodies of the invention having increased antibody-dependent cellular cytotoxicity include, but are not limited to, anti-human neuroblastoma monoclonal antibody (chCE7) produced by the methods of the invention, a chimeric anti-human renal cell carcinoma monoclonal antibody (ch-G250) produced by the methods of the invention, a humanized anti-HER2 monoclonal antibody (e.g., Trastuzumab (HERCEPTIN)) produced by the methods of the invention, a chimeric anti-human colon, lung, and breast carcinoma monoclonal antibody (ING-1) produced by the methods of the invention, a humanized anti-human 17-1A antigen monoclonal antibody (3622W94) produced by the methods of the invention, a humanized anti-human colorectal tumor antibody (A33) produced by the methods of the invention, an anti-human melanoma antibody (R24) directed against GD3 ganglioside produced by the methods of the invention, and a chimeric anti-human squamous-cell carcinoma
- the present invention relates to a method for increasing the ADCC activity of therapeutic antibodies. This is achieved by engineering the glycosylation pattern of the Fc region of such antibodies, in particular by maximizing the proportion of antibody molecules carrying bisected complex oligosaccharides and bisected hybrid oligosaccharides N-linked to the conserved glycosylation sites in their Fc regions.
- This strategy can be applied to increase Fc-mediated cellular cytotoxicity against undesirable cells mediated by any molecule carrying a region that is an equivalent to the Fc region of an immunoglobulin, not only by therapeutic antibodies, since the changes introduced by the engineering of glycosylation affect only the Fc region and therefore its interactions with the Fc receptors on the surface of effector cells involved in the ADCC mechanism.
- Fc-containing molecules to which the presently disclosed methods can be applied include, but are not limited to, (a) soluble fusion proteins made of a targeting protein domain fused to the N-terminus of an Fc-region (Chamov and Ashkenazi, Trends Biotech. 14: 52(1996) and (b) plasma membrane-anchored fusion proteins made of a type II transmembrane domain that localizes to the plasma membrane fused to the N-terminus of an Fc region (Stumble, P. F., Nature Biotech. 16: 1357 (1998)).
- the targeting domain directs binding of the fusion protein to undesirable cells such as cancer cells, i.e., in an analogous fashion to therapeutic antibodies.
- undesirable cells such as cancer cells, i.e., in an analogous fashion to therapeutic antibodies.
- the application of presently disclosed method to enhance the Fc-mediated cellular cytotoxic activity mediated by these molecules would therefore be identical to the method applied to therapeutic antibodies.
- the undesirable cells in the body have to express the gene encoding the fusion protein.
- This can be achieved either by gene therapy approaches, i.e., by transfecting the cells in vivo with a plasmid or viral vector that directs expression of the fusion protein-encoding gene to undesirable cells, or by implantation in the body of cells genetically engineered to express the fusion protein on their surface.
- the later cells would normally be implanted in the body inside a polymer capsule (encapsulated cell therapy) where they cannot be destroyed by an Fc-mediated cellular cytotoxicity mechanism. However should the capsule device fail and the escaping cells become undesirable, then they can be eliminated by Fc-mediated cellular cytotoxicity.
- the presently disclosed method would be applied either by incorporating into the gene therapy vector an additional gene expression cassette directing adequate or maximal expression levels of GnT III or by engineering the cells to be implanted to express adequate or maximal levels of GnT III.
- the aim of the disclosed method is to increase or maximize the proportion of surface-displayed Fc regions carrying bisected complex oligosaccharides and/or bisected hybrid oligosaccharides.
- VL and VH cDNA fragments were subcloned into pBluescriptIIKS(+), sequenced and directly joined by ligation to the human constant light (Ig ⁇ ) and heavy (IgG1) chain cDNAs, respectively, using unique restriction sites introduced at the variable and constant region junctions without altering the original amino acid residue sequence (Umana, P., et al., Nat Biotechnol. 17:176-180 (1999); Reff, M. E., et al., Blood 83:435-445 (1994)).
- IDEC-C2B8 in CHO cells expressing different levels of GnTIII.
- Establishment of two CHO cell lines, CHO-tet-GnTIII expressing different levels of GnTIII depending on the tetracycline concentration in the culture medium; and CHO-tTA, the parental cell line that does not express GnTIII has been described previously (Umana, P., et al., Nat Biotechnol. 17:176-180 (1999); Umana, P., et al., Biotechnol Bioeng. 65:542-549 (1999)).
- Each cell line was cotranfected with vectors pC2B8L, pC2B8H, and pZeoSV2(+) (for Zeocin resistance; Invitrogen, Leek, The Netherlands) using a calcium phosphate method.
- Zeocin resistant clones were transferred to a 96-well plate and assayed for IDEC-C2B8 production using an ELISA assay specific for the human constant region (4).
- Three IDEC-C2B8 samples were obtained from parallel cultures of a selected clone (CHO-tet-GnTIII-C2B8), differing only in the tetracycline concentration added to the medium (25, 50 and 2000 ng/mL respectively).
- CD20-positive cells SB cells; ATCC deposit no. ATCC CCL120
- CD20-negative cells HB cells; ATCC deposit no. ATCC CCL120.1
- SB cells CD20-positive cells
- HSB cells CD20-negative cells
- HBSSB bovine serum albumin fraction V
- HBSSB bovine serum albumin fraction V
- FITC-conjugated, anti-human Fc polyclonal antibody was used as a secondary antibody (SIGMA, St. Louis) for all samples. Cells were examined using a Leica fluorescence microscope (Wetzlar, Germany).
- Oligosaccharide profiling by MALDI/TOF-MS were derived from C2B8 antibody samples, MabTheraTM (European counterpart of Rituximab; kind gift from R. Stahel, Universit ⁇ dot over (a) ⁇ tspital, Switzerland), C2B8-25t, C2B8-50t, C2B8-2000t, and C2B8-nt, (100 ⁇ g each) as previously described (Umana, P., et al., Nat Biotechnol. 17:176-180 (1999)).
- the antibody samples were first treated with Arthrobacter ureafaciens sialidase (Oxford Glycosciences, Abingson, UK) to remove any sialic acid monosaccharide residues.
- Neutral N-linked oligosaccharides were then released from the desialylated antibody samples using peptide-N-glycosidase F (Oxford Glycosciences), purified using micro-columns, and analyzed by MALDI/TOF-MS in an Elite Voyager 400 spectrometer (Perseptive Biosystems, Farmingham, Mass.).
- PBMC Peripheral blood mononuclear cells
- IDEC-C2B8 Production of IDEC-C2B8 and verification of specific antigen binding.
- CHO-tet-GnTIII cells with stable, tetracycline-regulated expression of GnTIII and stable, constitutive expression of IDEC-C2B8, were established and scaled-up for production of a set of antibody samples.
- parallel cultures from the same clone were grown under three different tetracycline concentrations, 25, 50 and 2000 ng/ml. These levels of tetracycline had previously been shown to result in different levels of GnTIII and bisected oligosaccharides (Umana, P., et al., Nat Biotechnol.
- a C2B8-producing, control cell line that does not express GnTIII was also established and cultured under the same conditions as for the three parallel cultures of CHO-tet-GnTIII. After Protein A-affinity chromatography, mAb purity was estimated to be higher than 95% by SDS-PAGE and Coomassie-blue staining.
- Sample C2B 8-25t showed specific antigen binding by indirect immunofluorescence using CD20-positive and CD20-negative cells ( FIG. 1 ), indicating that the synthesized VL and VH gene fragments were functionally correct.
- Oligosaccharide profiling with MALDI/TOF-MS The glycosylation profile of each antibody sample was analyzed by MALDI/TOF-MS of the released, neutral oligosaccharide mix. In this technique, oligosaccharides of different mass appear as separate peaks in the spectrum and their proportions are quantitatively reflected by the relative peak heights (Harvey, D. J., Rapid Common Mass Spectrom. 7:614-619 (1993); Harvey, D. J., et al., Glycoconj J. 15:333-338 (1998)).
- Oligosaccharide structures were assigned to different peaks based on their expected molecular masses, previous structural data for oligosaccharides derived from IgG1 mAbs produced in the same host, and information on the N-linked oligosaccharide biosynthetic pathway.
- GnTIII expression levels i.e., tetracycline concentration
- C2B8-nt which are derived from hosts that do not express GnTIII
- FIGS. 2A and 2B did not carry bisected oligosaccharides
- bisected structures amounted up to approximately 35% of the oligosaccharides pool in sample C2B8-2000t, i.e, at a basal level of GnTIII expression.
- ADCC activity of IDEC-C2B8 glycosylated variants Different C2B8 mAb glycosylation variants were compared for ADCC activity, measured as in vitro lysis of CD20-positive SB cells.
- sample C2B8-50t carried approximately equal levels of bisected and non-bisected oligosaccharides, but did not mediate significantly higher target-cell lysis.
- sample C2B8-25t which contained up to 80% of bisected oligosaccharide structures, was significantly more active than the rest of the samples in the whole antibody concentration range. It reached the maximal level of ADCC activity of sample C2B8-nt at a 10-fold lower antibody concentration ( FIG. 4A ).
- Sample C2B8-25t also showed a significant increase in the maximal ADCC activity with respect to the control (50% vs. 30% lysis).
- SP2/0 mouse myeloma cells producing chG250 chimeric mAb were grown in standard cell culture medium supplemented with 1:100 (v/v) penicillin/streptomycin/antimycotic solution (SIGMA, Buchs, Switzerland). Cells were cultured at 37° C. in a 5% CO 2 humidified atmosphere in Tissue Culture Flasks. Medium was changed each 3-4 days. Cells were frozen in culture medium containing 10% DMSO.
- wt-chG250-SP2/O myeloma cells were transfected by electroporation with a vector for constitutive expression of GnTIII operatively linked via an IRES to a puromycin resistance gene. 24 hours before electroporation culture medium was changed and cells were seeded at 5 ⁇ 10 5 cells/ml. Seven million cells were centrifuged for 4 min at 1300 rpm at 4° C. Cells were washed with 3 mL new medium and centrifuged again. Cells were resuspended in a volume of 0.3-0.5 ml of reaction mix, containing 1.25% (v/v) DMSO and 20-30 ⁇ g DNA in culture medium.
- the electroporation mix was then transferred to a 0.4 cm cuvette and pulsed at low voltage (250-300 V) and high capacitance (960 ⁇ F) using Gene Pulser from Bio Rad. After electroporation cells were quickly transferred to 6 mL 1.25% (v/v) DMSO culture medium in a T25 culture flask and incubated at 37° C. Stable integrants were selected by applying 2 ⁇ g/mL puromycin to the medium two days after electroporation. After 2-3 weeks a stable, puromycin-resitant mixed population was obtained. Single-cell derived clones were obtained via FACS and were subsequently expanded and maintained under puromycin selection.
- Clones 2F1, 3D3, 4E6, 4E8, 4G3, 5G2, 5H12 and the wild type were seeded at 3 ⁇ 10 5 cells/mL in a total volume of 130 ml culture medium, and cultivated in single Triple-flasks. Cells used for seeding were all in full exponential growth phase, therefore cells were considered to be at the same growth state when the production batches started. Cells were cultivated for 4 days. Supernatants containing the antibody were collected in the late exponential growth phase to ensure reproducibility. The chG250 monoclonal antibody was purified in two chromatographic steps.
- Culture supernatants containing the chG250 monoclonal antibody derived from each batch were first purified using a HiTrap Protein A affinity chromatography. Protein A is highly specific for the human IgG F, region. Pooled samples from the protein A eluate were buffer exchanged to PBS by cation-exchange chromatography on a Resource S 1 ml column (Amersham Pharmacia Biotech). Final purity was judged to be higher than 95% from SDS-staining and Coomassie blue staining ( FIG. 6 ). The concentration of each sample was determined with a standard calibration curve using wild type antibody with known concentration.
- Oligosaccharide profiles were obtained by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI/TOF-MS), which accurately provides the molecular masses of the different oligosaccharide structures.
- MALDI/TOF-MS matrix-assisted laser desorption/ionization time of flight mass spectrometry
- This technique allows a quantitative analysis of proportions between different oligosaccharide structures within a mixture.
- Neutral oligosaccharides appeared predominantly as [M+Na + ] ions, however sometimes they were accompanied by-smaller [M+K +] ions, leading to an increase in mass of m/z of 16.
- the percentage of the structure appearing as potassium ion adducts depends on the content of the matrix and may thus vary between samples.
- a mixture of neutral N-linked oligosaccharides derived from each antibody preparation was analyzed using a 2,5-dehydrobenzoic acid (2,5-DHB) as matrix.
- 2,5-DHB 2,5-dehydrobenzoic acid
- Some of the peaks in the spectra were unequivocally assigned to specific oligosaccharide structures, because of known monosaccharide composition and unique mass. However, sometimes multiple structures could be assigned to a particular mass.
- MALDI enables the determination of the mass and cannot distinguish between isomers.
- Knowledge of the biosynthetic pathway and previous structural data enable, in most cases, the assignment of an oligosaccharide structure to a peak in the spectrum.
- GnTIII generated bisected F c -associated oligosaccharide structures of two types: complex or hybrid.
- Complex bisected oligosaccharides were unequivocally assigned to peaks at m/z 1543, 1689, 1705, 1851 and 1867 ([M+K + ] adduct).
- the increase in bisected oligosaccharides was accompanied by a concomitant reduction of peaks m/z 1486 and 1648, that correspond to nonbisected complex oligosaccharides.
- the main substrate of GnTIII m/z 1486) decreased dramatically.
- the percentage of the nonbisected complex oligosaccharide type assigned to peak at m/z 1648, had the lowest values for the clones expressing the highest GnTIII levels (clones 4E6, 4E8, 5G2 and 5H12). These two peaks decreased in favor of the accumulation of bisected complex and bisected hybrid type oligosaccharides ( FIGS. 7A-7D and 8 A- 8 D). The percentage of bisected complex oligosaccharides was higher for the samples derived from the clones expressing lower amounts of GnTIII. This is consistent with the fact that a higher GnTIII expression level probably shifts the biosynthetic flux to bisected hybrid structures, thereby decreasing the relative proportions of complex and complex bisected compound.
- Peaks m/z 1664, 1680, 1810 and 1826 can be assigned to either bisected hybrid type, to galactosylated complex oligosaccharides, or a mixture of them. Due to the fact that the wt-antibody preparation had a relatively low percentage of peak 1664, it was assumed that this peak, appearing in significant amounts in the antibody samples derived from the different clones, corresponded entirely to bisected hybrid structures ( FIGS. 7A-7D and 8 A- 8 D).
- the Calcein-AM retention method of measuring cytotoxicity measures the dye fluorescence remaining in the cells after incubation with the antibody.
- Four million G250 antigen-positive cells (target) were labelled with 10 ⁇ M Calcein-AM (Molecular Probes, Eugene, Oreg.) in 1.8 mL RPMI-1640 cell culture medium (GIBCO BRL, Basel, Switzerland) supplemented with 10% fetal calf serum for 30 min at 37° C. in a 5% CO 2 humidified atmosphere.
- the cells were washed twice in culture medium and resuspended in 12 mL AIMV serum free medium (GIBCO BRL, Basel, Switzerland).
- PBMC Peripheral blood mononuclear cells
- E:T ratio effector to target ratio
- 96-well plate was centrifuged at 700 ⁇ g for 5 min and the supernatants were discarded.
- the cell pellets were washed twice with Hank's balanced salt solution (HBSS) and lysed in 200 ⁇ L 0.05M sodium borate, pH 9, 0.1% Triton X-100. Retention of the fluorescent dye in the target cells was measured with a FLUOstar microplate reader (BMG LabTechnologies, Offenburg, Germany).
- the specific lysis was calculated relative to a total lysis control, resulting from exposure of the target cells to saponin (200 mg/mL in AIMV; SIGMA, Buchs, Switzerland) instead of exposure to antibody.
- G250 antigen-positive target cells were cultured with PBMCs with and without chG250 antibody samples at different concentrations.
- the cytotoxicity of unmodified chG250 antibody derived from the wild type cell line was compared with two antibody preparations derived from two cell lines (3D3, 5H12) expressing intermediate and high GnTIII levels, respectively (see FIG. 5 ).
- Unmodified chG250 antibody did not mediate significant ADCC activity over the entire concentration range used in the assay (the activity was not significantly different from background).
- Augmented ADCC activity (close to 20%, see FIG. 9 ) at 2 ⁇ g/mL was observed with the antibody sample derived from clone 3D3, which expressed intermediate GnTIII levels.
- the cytotoxic activity of this antibody samples did not grow at higher antibody concentrations.
- the antibody preparation derived from clone 5H12 showed a striking increase over samples 3D3 and unmodified antibody in its ability to mediate ADCC against target cells.
- the maximal ADCC activity of this antibody preparation was around 50% and was remarkable in mediating significant ADCC activity at 125-fold less concentrated when comparing with the unmodified control sample.
- Autoimmune thrombocytopenia in chronic graft-versus-host disease represents an instance of B-cell dysregulation leading to clinical disease.
- an anti-CD20 chimeric monoclonal antibody prepared by the methods of the present invention and having increased ADCC is administered to the subject as described in Ratanatharathorn, V. et al., Ann. Intern. Med. 133(4):275-79 (2000) (the entire contents of which is hereby incorporated by reference).
- a weekly infusion of the antibody 375 mg/m 2 is administered to the subject for 4 weeks.
- the antibody therapy produces a marked depletion of B cells in the peripheral blood and decreased levels of platelet-associated antibody.
- Immune-mediated, acquired pure red cell aplasia is a rare disorder frequently associated with other autoimmune phenomena
- an anti-CD20 chimeric monoclonal antibody prepared by the methods of the present invention and having increased ADCC is administered to the subject as described in Zecca, M. et al., Blood 12:3995-97 (1997) (the entire contents of which are hereby incorporated by reference).
- a subject with PRCA and autoimmune hemolytic anemia is given two doses of antibody, 375 mg/m 2 , per week.
- substitutive treatment with intravenous immunoglobulin is initiated. This treatment produces a marked depletion of B cells and a significant rise in reticulocyte count accompanied by increased hemoglobin levels.
Abstract
Description
- 1. Field of the Invention
- The present invention relates to the field of glycosylation engineering of proteins. More particularly, the present invention relates to glycosylation engineering to generate proteins with improved therapeutic properties, including antibodies with increased antibody-dependent cellular cytotoxicity.
- 2. Background Art
- Glycoproteins mediate many essential functions in human beings, other eukaryotic organisms, and some prokaryotes, including catalysis, signaling, cell-cell communication, and molecular recognition and association. They make up the majority of non-cytosolic proteins in eukaryotic organisms. (Lis et al., Eur. J. Biochem. 218:1-27 (1993)). Many glycoproteins have been exploited for therapeutic purposes, and during the last two decades, recombinant versions of naturally-occurring, secreted glycoproteins have been a major product of the biotechnology industry. Examples include erythropoietin (EPO), therapeutic monoclonal antibodies (therapeutic mAbs), tissue plasminogen activator (tPA), interferon-β, (IFN-β), granulocyte-macrophage colony stimulating factor (GM-CSF), and human chorionic gonadotrophin (hCG). (Cumming et al., Glycobiology 1:115-130 (1991)).
- The oligosaccharide component can significantly affect properties relevant to the efficacy of a therapeutic glycoprotein, including physical stability, resistance to protease attack, interactions with the immune system, pharmacokinetics, and specific biological activity. Such properties may depend not only on the presence or absence, but also on the specific structures, of oligosaccharides. Some generalizations between oligosaccharide structure and glycoprotein function can be made. For example, certain oligosaccharide structures mediate rapid clearance of the glycoprotein from the bloodstream through interactions with specific carbohydrate binding proteins, while others can be bound by antibodies and trigger undesired immune reactions. (Jenkins et al., Nature Biotechnol. 14:975-81 (1996)).
- Mammalian cells are the preferred hosts for production of therapeutic glycoproteins, due to their capability to glycosylate proteins in the most compatible form for human application. (Cumming et al., Glycobiology 1: 115-30 (1991); Jenkins et al., Nature Biotechnol. 14:975-81 (1996)). Bacteria very rarely glycosylate proteins, and like other types of common hosts, such as yeasts, filamentous fungi, insect and plant cells, yield glycosylation patterns associated with rapid clearance from the bloodstream, undesirable immune interactions, and in some specific cases, reduced biological activity. Among mammalian cells, Chinese hamster ovary (CHO) cells have been most commonly used during the last two decades. In addition to giving suitable glycosylation patterns, these cells allow consistent generation of genetically stable, highly productive clonal cell lines. They can be cultured to high densities in simple bioreactors using serum-free media, and permit the development of safe and reproducible bioprocesses. Other commonly used animal cells include baby hamster kidney (BHK) cells, NS0- and SP2/0-mouse myeloma cells. More recently, production from transgenic animals has also been tested (Jenkins et al., Nature Biotechnol. 14:975-81 (1996)).
- All antibodies contain carbohydrate structures at conserved positions in the heavy chain constant regions, with each isotype possessing a distinct array of N-linked carbohydrate structures, which variably affect protein assembly, secretion or functional activity. (Wright, A., and Morrison, S. L., Trends Biotech. 15:26-32 (1997)). The structure of the attached N-linked carbohydrate varies considerably, depending on the degree of processing, and can include high-mannose, multiply-branched as well as biantennary complex oligosaccharides. (Wright, A., and Morrison, S. L., Trends Biotech. 15:26-32 (1997)). Typically, there is heterogeneous processing of the core oligosaccharide structures attached at a particular glycosylation site such that even monoclonal antibodies exist as multiple glycoforms. Likewise, it has been shown that major differences in antibody glycosylation occur between cell lines, and even minor differences are seen for a given cell line grown under different culture conditions. (Lifely, M. R. et al., Glycobiology 5(8):813-22 (1995)).
- Unconjugated monoclonal antibodies (mAbs) can be useful medicines for the treatment of cancer, as demonstrated by the U.S. Food and Drug Administration's approval of Rituximab (Rituxan™; IDEC Pharmaceuticals, San Diego, Calif., and Genentech Inc., San Francisco, Calif.), for the treatment of CD20 positive B-cell, low-grade or follicular Non-Hodgkin's lymphoma, and Trastuzumab (Herceptin™; Genentech Inc,) for the treatment of advanced breast cancer (Grillo-Lopez, A.-J., et al., Semin. Oncol. 26:66-73 (1999); Goldenberg, M. M., Clin. Ther. 21:309-18 (1999)). The success of these products relies not only on their efficacy but also on their outstanding safety profiles (Grillo-Lopez, A.-J., et al., Semin. Oncol. 26:66-73 (1999); Goldenberg, M. M., Clin. Ther. 21:309-18 (1999)). In spite of the achievements of these two drugs, there is currently a large interest in obtaining higher specific antibody activity than what is typically afforded by unconjugated mAb therapy.
- One way to obtain large increases in potency, while maintaining a simple production process and potentially avoiding significant, undesirable side effects, is to enhance the natural, cell-mediated effector functions of mAbs by engineering their oligosaccharide component (Umaña, P. et al., Nature Biotechnol. 17:176-180 (1999)). IgG1 type antibodies, the most commonly used antibodies in cancer immunotherapy, are glycoproteins that have a conserved N-linked glycosylation site at Asn297 in each CH2 domain. The two complex bi-antennary oligosaccharides attached to Asn297 are buried between the CH2 domains, forming extensive contacts with the polypeptide backbone, and their presence is essential for the antibody to mediate effector functions such as antibody dependent cellular cytotoxicity (ADCC) (Lifely, M. R., et al., Glycobiology 5:813-822 (1995); Jefferis, R., et al., Immunol Rev. 163:59-76 (1998); Wright, A. and Morrison, S. L., Trends Biotechnol. 15:26-32 (1997)).
- The present inventors showed previously that overexpression in Chinese hamster ovary (CHO) cells of β(1,4)-N-acetylglucosaminyltransferase III (GnTIII), a glycosyltransferase catalyzing the formation of bisected oligosaccharides, significantly increases the in vitro ADCC activity of an anti-neuroblastoma chimeric monoclonal antibody (chCE7) produced by the engineered CHO cells. (See Umaña, P. et al., Nature Biotechnol. 17:176-180 (1999), International Publication No.
WO 99/54342, the entire contents of each of which are hereby incorporated by reference in their entirety). The antibody chCE7 belongs to a large class of unconjugated mAbs which have high tumor affinity and specificity, but have too little potency to be clinically useful when produced in standard industrial cell lines lacking the GnTIII enzyme (Umana, P., et al., Nature Biotechnol. 17:176-180 (1999)). That study was the first to show that large increases of maximal in vitro ADCC activity could be obtained by increasing the proportion of constant region (Fc)-associated, bisected oligosaccharides above the levels found in naturally occurring antibodies. To determine if this finding could be extrapolated to an unconjugated mAb, which already has significant ADCC activity in the absence of bisected oligosaccharides, the present inventors have applied this technology to Rituximab, the anti-CD20, IDEC-C2B8 chimeric antibody. The present inventors have likewise applied the technology to the unconjugated anti-cancer mAb chG250 - The present inventors have now generated new glycosylation variants of the anti-CD20 monoclonal antibody (mAb) IDEC-C2B8 (Rituximab) and the anti-cancer mAb chG250 using genetically engineered mAb-producing cell lines that overexpress N-acetylglucosaminyltransferase III (GnTIII; EC 2.1.4.144) in a tetracycline regulated fashion. GnTIII is required for the synthesis of bisected oligosaccharides, which are found at low to intermediate levels in naturally-occurring human antibodies but are missing in mAbs produced in standard industrial cell lines. The new glycosylated versions outperformed Mabthera™ (the version of Rixtuximab marketed in Europe) and mouse-myeloma derived chG250 in biological (ADCC) activity. For example, a ten-fold lower amount of the variant carrying the highest levels of bisected oligosaccharides was required to reach the maximal ADCC activity as Mabthera™. For chG250, the variant carrying the highest levels of bisected oligosaccharides mediated significant ADCC activity at a 125-fold lower concentration than that required to detect even low ADCC activity by the unmodified control chG250. A clear correlation was found between the level of GnTIII expression and ADCC activity.
- Accordingly, in one aspect the claimed invention is directed to a host cell engineered to produce a polypeptide having increased Fc-mediated cellular cytotoxicity by expression of at least one nucleic acid encoding β(1,4)-N-acetylglucosaminyltransferase III (GnT III), wherein the polypeptide produced by the host cell is selected from the group consisting of a whole antibody molecule, an antibody fragment, and a fusion protein which includes a region equivalent to the Fc region of an immunoglobulin, and wherein the GnT III is expressed in an amount sufficient to increase the proportion of said polypeptide carrying bisected hybrid oligosaccharides or galactosylated complex oligosaccharides or mixtures thereof in the Fc region relative to polypeptides carrying bisected complex oligosaccharides in the Fc region.
- In a preferred embodiment, the polypeptide is IgG or a fragment thereof, most preferably, IgG1 or a fragment thereof. In a further preferred embodiment, the polypeptide is a fusion protein that includes a region equivalent to the Fc region of a human IgG.
- In another aspect of the claimed invention, a nucleic acid molecule comprising at least one gene encoding GnTIII has been introduced into the host cell. In a preferred embodiment, at least one gene encoding GnTIII has been introduced into the host cell chromosome.
- Alternatively, the host cell has been engineered such that an endogenous GnT III gene is activated, for example, by insertion of a DNA element which increases gene expression into the host chromosome. In a preferred embodiment, the endogenous GnTIII has been activated by insertion of a promoter, an enhancer, a transcription factor binding site, a transposon, or a retroviral element or combinations thereof into the host cell chromosome. In another aspect, the host cell has been selected to carry a mutation triggering expression of an endogenous GnTIII. Preferably, the host cell is the CHO cell mutant lec 10.
- In a further preferred embodiment of the claimed invention, the at least one nucleic acid encoding a GnTIII is operably linked to a constitutive promoter element.
- In a further preferred embodiment, the host cell is a CHO cell, a BHK cell, a NS0 cell, a SP2/0 cell, or a hybridoma cell, a Y0 myeloma cell, a P3X63 mouse myeloma cell, a PER cell or a PER.C6 cell and said polypeptide is an anti-CD20 antibody. In another preferred embodiment, the host cell is a SP2/0 cell and the polypeptide is the monoclonal antibody chG250.
- In another aspect, the claimed invention is directed to a host cell that further comprises at least one transfected nucleic acid encoding an antibody molecule, an antibody fragment, or a fusion protein that includes a region equivalent to the Fc region of an immunoglobulin. In a preferred embodiment, the host cell comprises at least one transfected nucleic acid encoding an anti-CD20 antibody, the chimeric anti-human neuroblastoma monoclonal antibody chCE7, the chimeric anti-human renal cell carcinoma monoclonal antibody chG250, the chimeric anti-human colon, lung, and breast carcinoma monoclonal antibody ING-1, the humanized anti-human 17-1A antigen monoclonal antibody 3622W94, the humanized anti-human colorectal tumor antibody A33, the anti-human melanoma antibody directed against GD3 ganglioside R24, or the chimeric anti-human squamous-cell carcinoma monoclonal antibody SF-25, an anti-human EGFR antibody, an anti-human EGFRvIII antibody, an anti-human PSMA antibody, and anti-human PSCA antibody, an anti-human CD22 antibody, an anti-human CD30 antibody, an anti-human CD33 antibody, an anti-human CD38 antibody, an anti-human CD40 antibody, an anti-human CD45 antibody, an anti-human CD52 antibody, an anti-human CD138 antibody, an anti-human HLA-DR variant antibody, an anti-human EpCAM antibody, an anti-human CEA antibody, an anti-human MUC1 antibody, an anti-human MUC1 core protein antibody, an anti-human aberrantly glycosylated MUC1 antibody, an antibody against human fibronectin variants containing the ED-B domain, and an anti-human HER2/neu antibody.
- In another aspect, the claimed invention is directed to a method for producing a polypeptide in a host cell comprising culturing any of the above-described the host cells under conditions which permit the production of said polypeptide having increased Fc-mediated cellular cytotoxicity. In a preferred embodiment, the method further comprises isolating said polypeptide having increased Fc-mediated cellular cytotoxicity.
- In a further preferred embodiment, the host cell comprises at least one nucleic acid encoding a fusion protein comprising a region equivalent to a glycosylated Fc region of an immunoglobulin.
- In a preferred embodiment, the proportion of bisected oligosaccharides in the Fc region of said polypeptides is greater than 50%, more preferably, greater than 70%. In another embodiment, the proportion of bisected hybrid oligosaccharides or galactosylated complex oligosaccharides or mixtures thereof in the Fc region is greater than the proportion of bisected complex oligosaccharides in the Fc region of said polypeptide.
- In a preferred aspect of the claimed method, the polypeptide is an anti-CD20 antibody and the anti-CD20 antibodies produced by said host cell have a glycosylation profile, as analyzed by MALDI/TOF-MS, that is substantially equivalent to that shown in
FIG. 2E . - In another preferred aspect of the claimed method, the polypeptide is the chG250 monoclonal antibody and the chG250 antibodies produced by said host cell have a glycosylaton profile, as analyzed by MALDI/TOF-MS, that is substantially equivalent to that shown in
FIG. 7D . - In a further aspect, the claimed invention is directed to an antibody having increased antibody dependent cellular cytotoxicity (ADCC) produced by any of the methods described above. In preferred embodiments, the antibody is selected from the group consisting of an anti-CD20 antibody, chCE7, ch-G250, a humanized anti-HER2 monoclonal antibody, ING-1, 3622W94, SP-25, A33, and R24. Alternatively, the polypeptide can be an antibody fragment that includes a region equivalent to the Fc region of an immunoglobulin, having increased Fc-mediated cellular cytotoxicity produced by any of the methods described above.
- In a further aspect, the claimed invention is directed to a fusion protein that includes a region equivalent to the Fc region of an immunoglobulin and having increased Fc-mediated cellular cytotoxicity produced by any of the methods described above.
- In a further aspect, the claimed invention is directed to a pharmaceutical composition comprising the antibody, antibody fragment, or fusion protein of the invention and a pharmaceutically acceptable carrier.
- In a further aspect, the claimed invention is directed to a method for the treatment of cancer comprising administering a therapeutically effective amount of said pharmaceutical composition to a patient in need thereof.
- In a further aspect, the invention is directed to an improved method for treating an autoimmune disease produced in whole or in part by pathogenic autoantibodies based on B-cell depletion comprising administering a therapeutically effective amount of immunologically active antibody to a human subject in need thereof, the improvement comprising administering a therapeutically effective amount of an antibody having increased ADCC prepared as described above. In a preferred embodiment, the antibody is an anti-CD20 antibody. Examples of autoimmune diseases or disorders include, but are not limited to, immune-mediated thrombocytopenias, such as acute idiopathic thrombocytopenic purpurea and chronic idiopathic thrombocytopenic purpurea, dermatomyositis, Sydenham's chorea, lupus nephritis, rheumatic fever, polyglandular syndromes, Henoch-Schonlein purpura, post-streptococcal nephritis, erythema nodosum, Takayasu's arteritis, Addison's disease, erythema multiforme, polyarteritis nodosa, ankylosing spondylitis, Goodpasture's syndrome, thromboangitis ubiterans, primary biliary cirrhosis, Hashimoto's thyroiditis, thyrotoxicosis, chronic active hepatitis, polymyositis/dermatomyositis, polychondritis, pamphigus vulgaris, Wegener's granulomatosis, membranous-nephropathy, amyotrophic lateral sclerosis, tabes dorsalis, polymyaglia, pernicious anemia, rapidly progressive glomerulonephritis and fibrosing alveolitis, inflammatory responses such as inflammatory skin diseases including psoriasis and dermatitis (e.g. atopic dermatitis); systemic scleroderma and sclerosis; responses associated with inflammatory bowel disease (such as Crohn's disease and ulcerative colitis); respiratory distress syndrome (including adult respiratory distress syndrome; ARDS); dermatitis; meningitis; encephalitis; uveitis; colitis; glomerulonephritis; allergic conditions such as eczema and asthma and other conditions involving infiltration of T cells and chronic inflammatory responses; atherosclerosis; leukocyte adhesion deficiency; rheumatoid arthritis; systemic lupus erythematosus (SLE); diabetes mellitus (e.g. Type 1 diabetes mellitus or insulin dependent diabetes mellitus); multiple sclerosis; Reynaud's syndrome; autoimmune thyroiditis; allergic encephalomyelitis; Sjorgen's syndrome; juvenile onset diabetes; and immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes typically found in tuberculosis, sarcoidosis, polymyositis, granulomatosis and vasculitis; pernicious amenia (Addison's disease); diseases involving leukocyte diapedesis; central nervous system (CNS) inflammatory disorder, multiple organ injury syndrome; hemolytic anemia (including, but not limited to cryoglobinemia or Coombs positive anemia); myastheniagravis; antigen-antibody complex mediated diseases; anti-glomerular basement membrane disease; antiphospholipid syndrome; allergic neuritis; Graves' disease; Lambert-Eaton myasthenic syndrome; pemphigoid bullous; pemphigus; autoimmune polyendocrinopathies; Reiter's disease; stiff-man syndrome; Behcet disease; giant cell arteritis; immune complex nephritis; IgA nephropathy; IgM polyneuropathies; immune thrombocytopenic purpura (ITP) or autoimmune thrombocytopenia etc. In this aspect of the invention, the antibodies of the invention are used to deplete the blood of normal B-cells for an extended period.
-
FIG. 1 . Indirect immunofluorescence assay showing the reactivity of the antibody preparation C2B8-25t to CD20 positive SB cells. Negative controls, including the HSB CD20 negative cell line and cells treated only with the secondary FITC-conjugated anti-human Fc polyclonal antibody are not shown. -
FIG. 2A-2E . MALDI/TOF-MS spectra of the oligosaccharides derived from Mabthera™ (FIG. 2A ), C2B8-nt (FIG. 2B ), C2B8-2000t (FIG. 2C ), C2B8-50t (FIG. 2D ), and C2B8-25t (FIG. 2E ) antibody samples. Oligosaccharides appear as [M+Na+] and [M+K+] ions. Oligosaccharide appearing in the first two spectra were derived from cell cultures that do not express GnTIII, whereas oligosaccharides in C, D, and E were derived from a single cell line expressing GnTIII at different levels (i.e. tetracycline concentrations). -
FIGS. 3A and 3B . Illustration of a typical human IgG Fc-associated oligosaccharide structure (A) and partial N-linked glycosylation pathway (B). (FIG. 3A ) The core of the oligosaccharide is composed of three mannose (M) and two N-acetylglucosamine (Gn) monosaccharide residues attached to Asn297. Galactose (G), fucose (F), and bisecting N-acetylglucosamine (Gn, boxed) can be present or absent. Terminal N-acetylneuraminic acid maybe also present but it is not included in the figure. (FIG. 3B ) Partial N-linked glycosylation pathway leading to the formation of the major oligosaccharide classes (dotted frames). Bisecting N-acetylglucosamine is denoted as Gnb. Subscript numbers indicate how many monosaccharide residues are present in each oligosaccharide. Each structure appears together with its sodium-associated [M+Na+] mass. The mass of those structures that contain fucose (f) are also included. -
FIGS. 4A and 4B . ADCC activities of Rituximab glycosylation variants. The percentage of cytotoxicity was measured via lysis of 51Cr labeled CD20-positive SB cells by human lymphocytes (E:T ratio of 100:1) mediated by different mAb concentrations. (FIG. 4A ) Activity of C2B8 samples derived from a single cell line but produced at increasing GnTIII expression levels (i.e., decreasing tetracycline concentrations). The samples are C2B8-2000t, C2B8-50t, C2B8-25t, and C2B8-nt (control mAb derived from a clone that does not express GnTIII (FIG. 4B ) ADCC activity of C2B8-50t and C2B8-25t compared to Mabthera™. -
FIG. 5 . Western blot analysis of the seven GnTIII expressing clones and the wild type. 30 μg of each sample were loaded on a 8.75% SDS gel, transferred to a PVDF membrane and probed with the anti-c-myc monoclonal antibody (9E10). WT refers to wt-chG250-SP2/0 cells. -
FIG. 6 . SDS polyacrylamide gel electrophoresis of resolved purified antibody samples. -
FIG. 7A-7D . MALDI/TOF-MS spectra of neutral oligosaccharide mixtures from chG250 mAb samples produced by clones expressing different GnTIII levels and wt-chG250-SP2/0 cells: WT (FIG. 7A ), 2F1 (FIG. 7B ), 3D3 (FIG. 7C ), 4E6 (FIG. 7D ). -
FIG. 8A-8D . MALDI/TOF-MS spectra of neutral oligosaccharide mixtures from chG250 mAb samples produced by clones expressing different GnTIII levels: 4E8, (FIG. 8A ); 5G2, (FIG. 8B ); 4G3, (FIG. 8C ); 5H12, (FIG. 8D ). -
FIG. 9 . In vitro ADCC assay of antibody samples derived from control wt-chG250-SP2/-cells and GnTIII transected clones 3D3 and 5H12. - Terms are used herein as generally used in the art, unless otherwise defined as follows:
- As used herein, the term antibody is intended to include whole antibody molecules, antibody fragments, or fusionproteins that include a region equivalent to the Fc region of an immunoglobulin.
- As used herein, the term region equivalent to the Fc region of an immunoglobulin is intended to include naturally occurring allelic variants of the Fc region of an immunoglobulin as well as variants having alterations which produce substitutions, additions, or deletions but which do not decrease substantially the ability of the immunoglobulin to mediate antibody dependent cellular cytotoxicity. For example, one or more amino acids can be deleted from the N-terminus or C-terminus of the Fc region of an immunoglobulin without substantial loss of biological function. Such variants can be selected according to general rules known in the art so as to have minimal effect on activity. (See, e.g., Bowie, J. U. et al., Science 247:1306-10 (1990).
- As used herein, the term glycoprotein-modifying glycosyl transferase refers to β(1,4)-N-acetylglucosaminyltransferase III (GnTIII).
- As used herein, the terms engineer, engineered, engineering and glycosylation engineering are considered to include any manipulation of the glycosylation pattern of a naturally occurring polypeptide or fragment thereof. Glycosylation engineering includes metabolic engineering of the glycosylation machinery of a cell, including genetic manipulations of the oligosaccharide synthesis pathways to achieve altered glycosylation of glycoproteins expressed in cells. Furthermore, glycosylation engineering includes the effects of mutations and cell environment on glycosylation.
- As used herein, the term host cell covers any kind of cellular system which can be engineered to generate modified glycoforms of proteins, protein fragments, or peptides of interest, including antibodies and antibody fragments. Typically, the host cells have been manipulated to express optimized levels of GnT III. Host cells include cultured cells, e.g., mammalian cultured cells, such as CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells or hybridoma cells, yeast cells, and insect cells, to name only a few, but also cells comprised within a transgenic animal or cultured tissue.
- As used herein, the term Fc-mediated cellular cytotoxicity includes antibody-dependent cellular cytotoxicity and cellular cytotoxicity mediated by a soluble Fc-fusion protein containing a human Fc-region. It is an immune mechanism leading to the lysis of “antibody-targeted cells” by “human immune effector cells”, wherein:
-
- The “human immune effector cells” are a population of leukocytes that display Fc receptors on their surface through which they bind to the Fc-region of antibodies or of Fc-fusion proteins and perform effector functions. Such a population may include, but is not limited to, peripheral blood mononuclear cells (PBMC) and/or natural killer (NK) cells.
- The “antibody-targeted cells” are cells bound by the antibodies or Fc-fusion proteins. The antibodies or Fc fusion-proteins bind to target cells via the protein part N-terminal to the Fc region.
- As used herein, the term increased Fc-mediated cellular cytotoxicity is defined as either an increase in the number of “antibody-targeted cells” that are lysed in a given time, at a given concentration of antibody, or of Fc-fusion protein, in the medium surrounding the target cells, by the mechanism of Fc-mediated cellular cytotoxicity defined above, and/or a reduction in the concentration of antibody, or of Fc-fusion protein, in the medium surrounding the target cells, required to achieve the lysis of a given number of “antibody-targeted cells”, in a given time, by the mechanism of Fc-mediated cellular cytotoxicity. The increase in Fc-mediated cellular cytotoxicity is relative to the cellular cytotoxicity mediated by the same antibody, or Fc-fusion protein, produced by the same type of host cells, using the same standard production, purification, formulation and storage methods, which are known to those skilled in the art, but that has not been produced by host cells engineered to express the glycosyltransferase GnTIII by the methods described herein.
- By antibody having increased antibody dependent cellular cytotoxicity (ADCC) is meant an antibody having increased ADCC as determined by any suitable method known to those of ordinary skill in the art. One accepted in vitro ADCC assay is as follows:
-
- 1) the assay uses target cells that are known to express the target antigen recognized by the antigen-binding region of the antibody;
- 2) the assay uses human peripheral blood mononuclear cells (PBMCs), isolated from blood of a randomly chosen healthy donor, as effector cells;
- 3) the assay is carried out according to following protocol:
- i) the PBMCs are isolated using standard density centrifugation procedures and are suspended at 5×106 cells/ml in RPMI cell culture medium;
- ii) the target cells are grown by standard tissue culture methods, harvested from the exponential growth phase with a viability higher than 90%, washed in RPMI cell culture medium, labelled with 100 micro-Curies of 51Cr, washed twice with cell culture medium, and resuspended in cell culture medium at a density of 105 cells/ml;
- iii) 100 microliters of the final target cell suspension above are transferred to each well of a 96-well microtiter plate;
- iv) the antibody is serially-diluted from 4000 ng/ml to 0.04 ng/ml in cell culture medium and 50 microliters of the resulting antibody solutions are added to the target cells in the 96-well microtiter plate, testing in triplicate various antibody concentrations covering the whole concentration range above;
- v) for the maximum release (MR) controls, 3 additional wells in the plate containing the labelled target cells, receive 50 microliters of a 2% (V/V) aqueous solution of non-ionic detergent (Nonidet, Sigma, St. Louis), instead of the antibody solution (point iv above);
- vi) for the spontaneous release (SR) controls, 3 additional wells in the plate containing the labelled target cells, receive 50 microliters of RPMI cell culture medium instead of the antibody solution (point iv above);
- vii) the 96-well microtiter plate is then centrifuged at 50×g for 1 minute and incubated for 1 hour at 4° C.;
- viii) 50 microliters of the PBMC suspension (point i above) are added to each well to yield an effector:target cell ratio of 25:1 and the plates are placed in an incubator under 5% CO2 atmosphere at 37° C. for 4 hours;
- ix) the cell-free supernatant from each well is harvested and the experimentally released radioactivity (ER) is quantified using a gamma counter;
- x) the percentage of specific lysis is calculated for each antibody concentration according to the formula (ER−MR)/(MR−SR)×100, where ER is the average radioactivity quantified (see point ix above) for that antibody concentration, MR is the average radioactivity quantified (see point ix above) for the MR controls (see point v above), and SR is the average radioactivity quantified (see point ix above) for the SR controls (see point vi above);
- 4) “increased ADCC” is defined as either an increase in the maximum percentage of specific lysis observed within the antibody concentration range tested above, and/or a reduction in the concentration of antibody required to achieve one half of the maximum percentage of specific lysis observed within the antibody concentration range tested above. The increase in ADCC is relative to the ADCC, measured with the above assay, mediated by the same antibody, produced by the same type of host cells, using the same standard production, purification, formulation and storage methods, which are known to those skilled in the art, but that has not been produced by host cells engineered to overexpress the glycosyltransferase GnTIII.
- As used herein, the term anti-CD20 antibody is intended to mean an antibody which specifically recognizes a cell surface non-glycosylated phosphoprotein of 35,000 Daltons, typically designated as the human B lymphocyte restricted differentiation antigen Bp35, commonly referred to as CD20.
- Identification and Generation of Nucleic Acids Encoding a Protein for which Modification of the Glycosylation Pattern is Desired
- The present invention provides methods for the generation and use of host cell systems for the production of glycoforms of antibodies or antibody fragments or fusion proteins which include antibody fragments with increased antibody-dependent cellular cytotoxicity. Identification of target epitopes and generation of antibodies having potential therapeutic value, for which modification of the glycosylation pattern is desired, and isolation of their respective coding nucleic acid sequence is within the scope of the invention.
- Various procedures known in the art may be used for the production of antibodies to target epitopes of interest. Such antibodies include but are not limited to polyclonal, monoclonal, chimeric, single chain, Fab fragments and fragments produced by an Fab expression library. Such antibodies may be useful, e.g., as diagnostic or therapeutic agents. As therapeutic agents, neutralizing antibodies, i.e., those which compete for binding with a ligand, substrate or adapter molecule, are of especially preferred interest.
- For the production of antibodies, various host animals are immunized by injection with the target protein of interest including, but not limited to, rabbits, mice, rats, etc. Various adjuvants may be used to increase the immunological response, depending on the host species, including but not limited to Freund's (complete and incomplete), mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, saponin, oil emulsions, keyhole limpet hemocyanin, dinitrophenol, and potentially useful human adjuvants such as BCG (bacille Calmette-Guerin) and Corynebacterium parvum.
- Monoclonal antibodies to the target of interest may be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique originally described by Kohler and Milstein, Nature 256:495-97 (1975), the human B-cell hybridoma technique (Kosbor et al., Immunology Today 4:72 (1983); Cote et al., Proc. Natl. Acad. Sci. U.S.A. 80:2026-30 (1983) and the EBV-hybridoma technique (Cole et al., Monoclonal Antibodies and Cancer Therapy 77-96 (Alan R. Liss, Inc., 1985)). In addition, techniques developed for the production of “chimeric antibodies” (Morrison et al., Proc. Natl. Acad. Sci. U.S.A. 81:6851-55 (1984); Neuberger et al., Nature 312:604-08 (1984); Takeda et al., Nature 314:452-54 (1985) by splicing the genes from a mouse antibody molecule of appropriate antigen specificity together with genes from a human antibody molecule of appropriate biological activity can be used. Alternatively, techniques described for the production of single chain antibodies (U.S. Pat. No. 4,946,778) can be adapted to produce single chain antibodies having a desired specificity.
- Antibody fragments which contain specific binding sites of the target protein of interest may be generated by known techniques. For example, such fragments include, but are not limited to, F(ab′)2 fragments which can be produced by pepsin digestion of the antibody molecule and the Fab fragments which can be generated by reducing the disulfide bridges of the F(ab′)2 fragments. Alternatively, Fab expression libraries may be constructed (Huse et al., Science 246:1275-81 (1989) to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity to the target protein of interest.
- Once an antibody or antibody fragment has been identified for which modification in the glycosylation pattern are desired, the coding nucleic acid sequence is identified and isolated using techniques well known in the art.
- a. Generation of Cell Lines for the Production of Proteins with Altered Glycosylation Pattern
- The present invention provides host cell expression systems for the generation of proteins having modified glycosylation patterns. In particular, the present invention provides host cell systems for the generation of glycoforms of proteins having an improved therapeutic value. Therefore, the invention provides host cell expression systems selected or engineered to increase the expression of a glycoprotein-modifying glycosyltransferase, namely β(1,4)-N-acetylglucosaminyltransferase III (GnTIII). Specifically, such host cell expression systems may be engineered to comprise a recombinant nucleic acid molecule encoding GnTIII, operatively linked to a constitutive or regulated promoter system. Alternatively, host cell expression systems may be employed that naturally produce, are induced to produce, and/or are selected to produce GnTIII.
- In one specific embodiment, the present invention provides a host cell that has been engineered to express at least one nucleic acid encoding GnTIII. In one aspect, the host cell is transformed or transfected with a nucleic acid molecule comprising at least one gene encoding GnTIII. In an alternate aspect, the host cell has been engineered and/or selected in such way that endogenous GnTIII is activated. For example, the host cell may be selected to carry a mutation triggering expression of endogenous GnTIII. In one specific embodiment, the host cell is a CHO lec10 mutant. Alternatively, the host cell may be engineered such that endogenous GnTIII is activated. In again another alternative, the host cell is engineered such that endogenous GnTIII has been activated by insertion of a constitutive promoter element, a transposon, or a retroviral element into the host cell chromosome.
- Generally, any type of cultured cell line can be used as a background to engineer the host cell lines of the present invention. In a preferred embodiment, CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells or hybridoma cells, yeast cells, or insect cells are used as the background cell line to generate the engineered host cells of the invention.
- The invention is contemplated to encompass any engineered host cells expressing GnTIII as defined herein.
- One or several nucleic acids encoding GnTIII may be expressed under the control of a constitutive promoter or, alternately, a regulated expression system. Suitable regulated expression systems include, but are not limited to, a tetracycline-regulated expression system, an ecdysone-inducible expression system, a lac-switch expression system, a glucocorticoid-inducible expression system, a temperature-inducible promoter system, and a metallothionein metal-inducible expression system. If several different nucleic acids encoding GnTIII are comprised within the host cell system, some of them may be expressed under the control of a constitutive promoter, while others are expressed under the control of a regulated promoter. The maximal expression level is considered to be the highest possible level of stable GnTIII expression that does not have a significant adverse effect on cell growth rate, and will be determined using routine experimentation. Expression levels are determined by methods generally known in the art, including Western blot analysis using a GnTIII specific antibody, Northern blot analysis using a GnTIII specific nucleic acid probe, or measurement of enzymatic activity. Alternatively, a lectin may be employed which binds to biosynthetic products of the GnTIII, for example, E4-PHA lectin. In a further alternative, the nucleic acid may be operatively linked to a reporter gene; the expression levels of the GnTIII are determined by measuring a signal correlated with the expression level of the reporter gene. The reporter gene may transcribed together with the nucleic acid(s) encoding said GnTIII as a single mRNA molecule; their respective coding sequences may be linked either by an internal ribosome entry site (IRES) or by a cap-independent translation enhancer (CITE). The reporter gene may be translated together with at least one nucleic acid encoding said GnTIII such that a single polypeptide chain is formed. The nucleic acid encoding the GnTIII may be operatively linked to the reporter gene under the control of a single promoter, such that the nucleic acid encoding the GnTIII and the reporter gene are transcribed into an RNA molecule which is alternatively spliced into two separate messenger RNA (mRNA) molecules; one of the resulting mRNAs is translated into said reporter protein, and the other is translated into said GnTIII.
- If several different nucleic acids encoding GnTIII are expressed, they may be arranged in such way that they are transcribed as one or as several mRNA molecules. If they are transcribed as a single mRNA molecule, their respective coding sequences may be linked either by an internal ribosome entry site (IRES) or by a cap-independent translation enhancer (CITE). They may be transcribed from a single promoter into an RNA molecule which is alternatively spliced into several separate messenger RNA (mRNA) molecules, which then are each translated into their respective encoded GnTIII.
- In other embodiments, the present invention provides host cell expression systems for the generation of therapeutic antibodies, having an increased antibody-dependent cellular cytotoxicity, and cells which display the IgG Fc region on the surface to promote Fc-mediated cytotoxicity. Generally, the host cell expression systems have been engineered and/or selected to express nucleic acids encoding the antibody for which the production of altered glycoforms is desired, along with at least one nucleic acid encoding GnTIII. In one embodiment, the host cell system is transfected with at least one gene encoding GnTIII. Typically, the transfected cells are selected to identify and isolate clones that stably express the GnTIII. In another embodiment, the host cell has been selected for expression of endogenous GnTIII. For example, cells may be selected carrying mutations which trigger expression of otherwise silent GnTIII. For example, CHO cells are known to carry a silent GnT III gene that is active in certain mutants, e.g., in the mutant Lec10. Furthermore, methods known in the art may be used to activate silent GnTIII, including the insertion of a regulated or constitutive promoter, the use of transposons, retroviral elements, etc. Also the use of gene knockout technologies or the use of ribozyme methods may be used to tailor the host cell's GnTIII expression level, and is therefore within the scope of the invention.
- Any type of cultured cell line can be used as background to engineer the host cell lines of the present invention. In a preferred embodiment, CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells or hybridoma cells, yeast cell, or insect cells may be used. Typically, such cell lines are engineered to further comprise at least one transfected nucleic acid encoding a whole antibody molecule, an antibody fragment, or a fusion protein that includes a region equivalent to the Fc region of an immunoglobulin. In an alternative embodiment, a hybridoma cell line expressing a particular antibody of interest is used as background cell line to generate the engineered host cells of the invention.
- Typically, at least one nucleic acid in the host cell system encodes GnT III.
- One or several nucleic acids encoding GnTIII may be expressed under the control of a constitutive promoter, or alternately, a regulated expression system. Suitable regulated expression systems include, but are not limited to, a tetracycline-regulated expression system, an ecdysone-inducible expression system, a lac-switch expression system, a glucocorticoid-inducible expression system, a temperature-inducible promoter system, and a metallothionein metal-inducible expression system. If several different nucleic acids encoding GnTIII are comprised within the host cell system, some of them may be expressed under the control of a constitutive promoter, while others are expressed under the control of a regulated promoter. The maximal expression level is considered to be the highest possible level of stable GnTIII expression that does not have a significant adverse effect on cell growth rate, and will be determined using routine experimentation. Expression levels are determined by methods generally known in the art, including Western blot analysis using a GnTIII specific antibody, Northern blot analysis using a GnTIII specific nucleic acid probe, or measurement of GnTIII enzymatic activity. Alternatively, a lectin may be employed which binds to biosynthetic products of GnTIII, for example, E4-PHA lectin. In a further alternative, the nucleic acid may be operatively linked to a reporter gene; the expression levels of the glycoprotein-modifying glycosyl transferase are determined by measuring a signal correlated with the expression level of the reporter gene. The reporter gene may transcribed together with the nucleic acid(s) encoding said glycoprotein-modifying glycosyl transferase as a single mRNA molecule; their respective coding sequences may be linked either by an internal ribosome entry site (IRES) or by a cap-independent translation enhancer (CITE). The reporter gene may be translated together with at least one nucleic acid encoding GnTIII such that a single polypeptide chain is formed. The nucleic acid encoding the GnTIII may be operatively linked to the reporter gene under the control of a single promoter, such that the nucleic acid encoding the GnTIII and the reporter gene are transcribed into an RNA molecule which is alternatively spliced into two separate messenger RNA (mRNA) molecules; one of the resulting mRNAs is translated into said reporter protein, and the other is translated into said GnTIII.
- If several different nucleic acids encoding a GnTIII are expressed, they may be arranged in such way that they are transcribed as one or as several mRNA molecules. If they are transcribed as single mRNA molecule, their respective coding sequences may be linked either by an internal ribosome entry site (IRES) or by a cap-independent translation enhancer (CITE). They may be transcribed from a single promoter into an RNA molecule which is alternatively spliced into several separate messenger RNA (mRNA) molecules, which then are each translated into their respective encoded GnTIII.
- i. Expression Systems Methods which are well known to those skilled in the art can be used to construct expression vectors containing the coding sequence of the protein of interest and the coding sequence of the GnTIII and appropriate transcriptional/translational control signals. These methods include in vitro recombinant DNA techniques, synthetic techniques and in vivo recombination/genetic recombination. See, for example, the techniques described in Maniatis et al., Molecular Cloning A Laboratory Manual, Cold Spring Harbor Laboratory, N.Y. (1989) and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, N.Y (1989).
- A variety of host-expression vector systems may be utilized to express the coding sequence of the protein of interest and the coding sequence of the GnTIII. Preferably, mammalian cells are used as host cell systems transfected with recombinant plasmid DNA or cosmid DNA expression vectors containing the coding sequence of the protein of interest and the coding sequence of the GnTIII. Most preferably, CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER. C6 cells or hybridoma cells, yeast cells, or insect cells are used as host cell system. In alternate embodiments, other eukaryotic host cell systems may be contemplated, including, yeast cells transformed with recombinant yeast expression vectors containing the coding sequence of the protein of interest and the coding sequence of the GnTIII; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing the coding sequence of the protein of interest and the coding sequence of the GnTIII; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing the coding sequence of the protein of interest and the coding sequence of the GnTIII; or animal cell systems infected with recombinant virus expression vectors (e.g., adenovirus, vaccinia virus) including cell lines engineered to contain multiple copies of the DNA encoding the protein of interest and the coding sequence of the GnTIII either stably amplified (CHO/dhfr) or unstably amplified in double-minute chromosomes (e.g., murine cell lines).
- For the methods of this invention, stable expression is generally preferred to transient expression because it typically achieves more reproducible results and also is more amenable to large scale production. Rather than using expression vectors which contain viral origins of replication, host cells can be transformed with the respective coding nucleic acids controlled by appropriate expression control elements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.), and a selectable marker. Following the introduction of foreign DNA, engineered cells may be allowed to grow for 1-2 days in an enriched media, and then are switched to a selective media. The selectable marker in the recombinant plasmid confers resistance to the selection and allows selection of cells which have stably integrated the plasmid into their chromosomes and grow to form foci which in turn can be cloned and expanded into cell lines.
- A number of selection systems may be used, including, but not limited to, the herpes simplex virus thymidine kinase (Wigler et al., Cell 11:223 (1977)), hypoxanthine-guanine phosphoribosyltransferase (Szybalska & Szybalski, Proc. Natl. Acad. Sci. USA 48:2026 (1962)), and adenine phosphoribosyltransferase (Lowy et al., Cell 22:817 (1980)) genes, which can be employed in tk−, hgprt− or aprt− cells, respectively. Also, antimetabolite resistance can be used as the basis of selection for dhfr, which confers resistance to methotrexate (Wigler et al., Natl. Acad. Sci. USA 77:3567 (1989); O'Hare et al., Proc. Natl. Acad. Sci. USA 78:1527 (1981)); gpt, which confers resistance to mycophenolic acid (Mulligan & Berg, Proc. Natl. Acad. Sci. USA 78:2072 (1981)); neo, which confers resistance to the aminoglycoside G-418 (Colberre-Garapin et al., J. Mol. Biol. 150:1 (1981)); and hygro, which confers resistance to hygromycin (Santerre et al., Gene 30:147 (1984) genes. Recently, additional selectable genes have been described, namely trpB, which allows cells to utilize indole in place of tryptophan; hisD, which allows cells to utilize histinol in place of histidine (Hartman & Mulligan, Proc. Natl. Acad. Sci. USA 85:8047 (1988)); the glutamine synthase system; and ODC (ornithine decarboxylase) which confers resistance to the ornithine decarboxylase inhibitor, 2-(difluoromethyl)-DL-ornithine, DFMO (McConlogue, in: Current Communications in Molecular Biology, Cold Spring Harbor Laboratory ed. (1987)).
- ii. Identification Of Transfectants or Transformants that Express the Protein Having a Modified Glycosylation Pattern
- The host cells which contain the coding sequence and which express the biologically active gene products may be identified by at least four general approaches; (a) DNA-DNA or DNA-RNA hybridization; (b) the presence or absence of “marker” gene functions; (c) assessing the level of transcription as measured by the expression of the respective mRNA transcripts in the host cell; and (d) detection of the gene product as measured by immunoassay or by its biological activity.
- In the first approach, the presence of the coding sequence of the protein of interest and the coding sequence of the GnTIII inserted in the expression vector can be detected by DNA-DNA or DNA-RNA hybridization using probes comprising nucleotide sequences that are homologous to the respective coding sequences, respectively, or portions or derivatives thereof.
- In the second approach, the recombinant expression vector/host system can be identified and selected based upon the presence or absence of certain “marker” gene functions (e.g., thymidine kinase activity, resistance to antibiotics, resistance to methotrexate, transformation phenotype, occlusion body formation in baculovirus, etc.). For example, if the coding sequence of the protein of interest and the coding sequence of the GnTIII are inserted within a marker gene sequence of the vector, recombinants containing the respective coding sequences can be identified by the absence of the marker gene function. Alternatively, a marker gene can be placed in tandem with the coding sequences under the control of the same or different promoter used to control the expression of the coding sequences. Expression of the marker in response to induction or selection indicates expression of the coding sequence of the protein of interest and the coding sequence of the GnTIII.
- In the third approach, transcriptional activity for the coding region of the protein of interest and the coding sequence of the GnTIII can be assessed by hybridization assays. For example, RNA can be isolated and analyzed by Northern blot using a probe homologous to the coding sequences of the protein of interest and the coding sequence of the GnTIII or particular portions thereof. Alternatively, total nucleic acids of the host cell may be extracted and assayed for hybridization to such probes.
- In the fourth approach, the expression of the protein products of the protein of interest and the coding sequence of the GnTIII can be assessed immunologically, for example by Western blots, immunoassays such as radioimmuno-precipitation, enzyme-linked immunoassays and the like. The ultimate test of the success of the expression system, however, involves the detection of the biologically active gene products.
- b. Generation and Use of Proteins and Protein Fragments Having Altered Glycosylation Patterns
- i. Generation and Use of Antibodies Having Increased Antibody-Dependent Cellular Cytotoxicity
- In preferred embodiments, the present invention provides glycoforms of antibodies and antibody fragments having increased antibody-dependent cellular cytotoxicity.
- Clinical trials of unconjugated monoclonal antibodies (mAbs) for the treatment of some types of cancer have recently yielded encouraging results. Dillman, Cancer Biother. & Radiopharm. 12:223-25 (1997); Deo et al., Immunology Today 18:127 (1997). A chimeric, unconjugated IgG1 has been approved for low-grade or follicular B-cell non-Hodgkin's lymphoma Dillman, Cancer Biother. & Radiopharm. 12:223-25 (1997), while another unconjugated mAb, a humanized IgG1 targeting solid breast tumors, has also been showing promising results in phase III clinical trials. Deo et al., Immunology Today 18:127 (1997). The antigens of these two mAbs are highly expressed in their respective tumor cells and the antibodies mediate potent tumor destruction by effector cells in vitro and in vivo. In contrast, many other unconjugated mAbs with fine tumor specificities cannot trigger effector functions of sufficient potency to be clinically useful. Frost et al., Cancer 80:317-33 (1997); Surfus et al., J. Immunother. 19:184-91 (1996). For some of these weaker mAbs, adjunct cytokine therapy is currently being tested. Addition of cytokines can stimulate antibody-dependent cellular cytotoxicity (ADCC) by increasing the activity and number of circulating lymphocytes. Frost et al., Cancer 80:317-33 (1997); Surfus et al., J. Immunother. 19:1.84-91 (1996). ADCC, a lytic attack on antibody-targeted cells, is triggered upon binding of leukocyte receptors to the constant region (Fc) of antibodies. Deo et al., Immunology Today 18:127 (1997).
- A different, but complementary, approach to increase ADCC activity of unconjugated IgG1s is to engineer the Fc region of the antibody to increase its affinity for the lymphocyte receptors (FcγRs). Protein engineering studies have shown that FcγRs interact with the lower hinge region of the IgG CH2 domain. Lund et al., J. Immunol. 157:4963-69 (1996). However, FcγR binding also requires the presence of oligosaccharides covalently attached at the conserved Asn 297 in the CH2 region. Lund et al., J. Immunol. 157:4963-69 (1996); Wright and Morrison, Trends Biotech. 15:26-31 (1997), suggesting that either oligosaccharide and polypeptide both directly contribute to the interaction site or that the oligosaccharide is required to maintain an active CH2 polypeptide conformation. Modification of the oligosaccharide structure can therefore be explored as a means to increase the affinity of the interaction.
- An IgG molecule carries two N-linked oligosaccharides in its Fc region, one on each heavy chain. As any glycoprotein, an antibody is produced as a population of glycoforms which share the same polypeptide backbone but have different oligosaccharides attached to the glycosylation sites. The oligosaccharides normally found in the Fc region of serum IgG are of complex bi-antennary type (Wormald et al., Biochemistry 36:130-38 (1997), with low level of terminal sialic acid and bisecting N-acetylglucosamine (GlcNAc), and a variable degree of terminal galactosylation and core fucosylation. Some studies suggest that the minimal carbohydrate structure required for FcγR binding lies within the oligosaccharide core. Lund et al., J. Immunol. 157:4963-69 (1996) The removal of terminal galactoses results in approximately a two-fold reduction in ADCC activity, indicating a role for these residues in FcγR receptor binding. Lund et al, J. Immunol. 157:4963-69 (1996)
- The mouse or hamster-derived cell lines used in industry and academia for production of unconjugated therapeutic mAbs normally attach the required oligosaccharide determinants to Fc sites. IgGs expressed in these cell lines lack, however, the bisecting GlcNAc found in low amounts in serum IgGs. Lifely et al., Glycobiology 318:813-22 (1995). In contrast, it was recently observed that a rat myeloma-produced, humanized IgG1 (CAMPATH-1H) carried a bisecting GlcNAc in some of its glycoforms. Lifely et al., Glycobiology 318:813-22 (1995). The rat cell-derived antibody reached a similar in vitro ADCC activity as CAMPATH-1H antibodies produced in standard cell lines, but at significantly lower antibody concentrations.
- The CAMPATH antigen is normally present at high levels on lymphoma cells, and this chimeric mAb has high ADCC activity in the absence of a bisecting GlcNAc. Lifely et al., Glycobiology 318:813-22 (1995). In the N-linked glycosylation pathway, a bisecting GlcNAc is added by the enzyme β(1,4)-N-acetylglucosaminyltransferase III (GnT III). Schachter, Biochem. Cell Biol. 64:163-81 (1986).
- The present inventors used a single antibody-producing CHO cell line, that was previously engineered to express, in an externally-regulated fashion, different levels of a cloned GnT III gene. This approach established for the first time a rigorous correlation between expression of GnTIII and the ADCC activity of the modified antibody.
- The present inventors previously showed that C2B8 antibody modified according to the disclosed method had an about sixteen-fold higher ADCC activity than the standard, unmodified C2B8 antibody produced under identical cell culture and purification conditions. Briefly, a C2B8 antibody sample expressed in CHO-tTA-C2B8 cells that do not have GnT III expression showed a cytotoxic activity of about 31% (at 1 μg/ml antibody concentration), measured as in vitro lysis of SB cells (CD20+) by human lymphocytes. In contrast, C2B8 antibody derived from a CHO cell culture expressing GnT III at a basal, largely repressed level showed at 1 μg/ml antibody concentration a 33% increase in ADCC activity against the control at the same antibody concentration. Moreover, increasing the expression of GnT III produced a large increase of almost 80% in the maximal ADCC activity (at 1 μg/ml antibody concentration) compared to the control at the same antibody concentration. (See International Publication No. WO 99/54342, the entire contents of which are hereby incorporated by reference.)
- Further antibodies of the invention having increased antibody-dependent cellular cytotoxicity include, but are not limited to, anti-human neuroblastoma monoclonal antibody (chCE7) produced by the methods of the invention, a chimeric anti-human renal cell carcinoma monoclonal antibody (ch-G250) produced by the methods of the invention, a humanized anti-HER2 monoclonal antibody (e.g., Trastuzumab (HERCEPTIN)) produced by the methods of the invention, a chimeric anti-human colon, lung, and breast carcinoma monoclonal antibody (ING-1) produced by the methods of the invention, a humanized anti-human 17-1A antigen monoclonal antibody (3622W94) produced by the methods of the invention, a humanized anti-human colorectal tumor antibody (A33) produced by the methods of the invention, an anti-human melanoma antibody (R24) directed against GD3 ganglioside produced by the methods of the invention, and a chimeric anti-human squamous-cell carcinoma monoclonal antibody (SF-25) produced by the methods of the invention, an anti-human small cell lung carcinoma monoclonal antibody (BEC2, ImClone Systems, Merck KgaA) produced by the methods of the invention, an anti-human non-Hodgkin's lymphoma monoclonal antibody (Bexxar (tositumomab, Coulter Pharmaceuticals), Oncolym (Techniclone, Alpha Therapeutic)) produced by the methods of the invention, an anti-human squamous cell head and neck carcinoma monoclonal antibody (C225, ImClone Systems) prepared by the methods of the invention, an anti-human rectal and colon carcinoma monoclonal antibody (Panorex (edrecolomab), Centocor, Glaxo Wellcome) prepared by the methods of the invention, an anti-human ovarian carcinoma monoclonal antibody (Theragyn, Antisoma) produced by the methods of the invention, an anti-human acute myelogenous leukemia carcinoma monoclonal antibody (SmartM195, Protein Design Labs, Kanebo) produced by the methods of the invention, an anti-human malignant glioma monoclonal antibody (Cotara, Techniclone, Cambridge Antibody Technology) produced by the methods of the invention, an anti-human B cell non-Hodgkins lymphoma monoclonal antibody (IDEC-Y2B8, IDEC Pharmaceuticals) produced by the methods of the invention, an anti-human solid tumors monoclonal antibody (CEA-Cide, Immunomedics) produced by the methods of the invention, an anti-human colorectal carcinoma monoclonal antibody (Iodine 131-MN-14, Immunomedics) produced by the methods of the invention, an anti-human ovary, kidney, breast, and prostate carcinoma monoclonal antibody (MDX-210, Medarex, Novartis) produced by the methods of the invention, an anti-human colorectal and pancreas carcinoma monoclonal antibody (TTMA, Pharmacie & Upjohn) produced by the methods of the invention, an anti-human TAG-72 expressing carcinoma monoclonal antibody (MDX-220, Medarex) produced by the methods of the invention, an anti-human EGFr-expressing carcinoma monoclonal antibody (MDX-447) produced by the methods of the invention, Anti-VEGF monoclonal antibody (Genentech) produced by the methods of the invention, an anti-human breast, lung, prostate and pancreas carcinoma and malignant melanoma monoclonal antibody (BrevaRex, AltaRex) produced by the methods of the invention, and an anti-human acute myelogenous leukemia monoclonal antibody (Monoclonal Antibody Conjugate, Immunex) produced by the methods of the invention. In addition, the invention is directed to antibody fragment and fusion proteins comprising a region that is equivalent to the Fc region of immunoglobulins.
- ii. Generation and Use of Fusion Proteins Comprising a Region Equivalent to an Fc Region of an Immunoglobulin that Promote Fc-Mediated Cytotoxicity
- As discussed above, the present invention relates to a method for increasing the ADCC activity of therapeutic antibodies. This is achieved by engineering the glycosylation pattern of the Fc region of such antibodies, in particular by maximizing the proportion of antibody molecules carrying bisected complex oligosaccharides and bisected hybrid oligosaccharides N-linked to the conserved glycosylation sites in their Fc regions. This strategy can be applied to increase Fc-mediated cellular cytotoxicity against undesirable cells mediated by any molecule carrying a region that is an equivalent to the Fc region of an immunoglobulin, not only by therapeutic antibodies, since the changes introduced by the engineering of glycosylation affect only the Fc region and therefore its interactions with the Fc receptors on the surface of effector cells involved in the ADCC mechanism. Fc-containing molecules to which the presently disclosed methods can be applied include, but are not limited to, (a) soluble fusion proteins made of a targeting protein domain fused to the N-terminus of an Fc-region (Chamov and Ashkenazi, Trends Biotech. 14: 52(1996) and (b) plasma membrane-anchored fusion proteins made of a type II transmembrane domain that localizes to the plasma membrane fused to the N-terminus of an Fc region (Stabila, P. F., Nature Biotech. 16: 1357 (1998)).
- In the case of soluble fusion proteins (a) the targeting domain directs binding of the fusion protein to undesirable cells such as cancer cells, i.e., in an analogous fashion to therapeutic antibodies. The application of presently disclosed method to enhance the Fc-mediated cellular cytotoxic activity mediated by these molecules would therefore be identical to the method applied to therapeutic antibodies.
- In the case of membrane-anchored fusion proteins (b) the undesirable cells in the body have to express the gene encoding the fusion protein. This can be achieved either by gene therapy approaches, i.e., by transfecting the cells in vivo with a plasmid or viral vector that directs expression of the fusion protein-encoding gene to undesirable cells, or by implantation in the body of cells genetically engineered to express the fusion protein on their surface. The later cells would normally be implanted in the body inside a polymer capsule (encapsulated cell therapy) where they cannot be destroyed by an Fc-mediated cellular cytotoxicity mechanism. However should the capsule device fail and the escaping cells become undesirable, then they can be eliminated by Fc-mediated cellular cytotoxicity. Stabila et al., Nature Biotech. 16: 1357 (1998). In this case, the presently disclosed method would be applied either by incorporating into the gene therapy vector an additional gene expression cassette directing adequate or maximal expression levels of GnT III or by engineering the cells to be implanted to express adequate or maximal levels of GnT III. In both cases, the aim of the disclosed method is to increase or maximize the proportion of surface-displayed Fc regions carrying bisected complex oligosaccharides and/or bisected hybrid oligosaccharides.
- The examples below explain the invention in more detail. The following preparations and examples are given to enable those skilled in the art to more clearly understand and to practice the present invention. The present invention, however, is not limited in scope by the exemplified embodiments, which are intended as illustrations of single aspects of the invention only, and methods which are functionally equivalent are within the scope of the invention. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.
- Synthesis of VH and VL coding regions of IDEC-C2B8 and construction of mammalian expression vectors. cDNAs encoding the VH and VL regions of IDEC-C2B8 antibody were assembled from a set of overlapping single-stranded oligonucleotides in a one-step process using PCR (Kobayashi, N., et al., Biotechniques 23:500-503(1997)). The original sequence data coding for IDEC-C2B8 VL and VH were obtained from a published international patent application (International Publication Number: WO 94/11026). Assembled VL and VH cDNA fragments were subcloned into pBluescriptIIKS(+), sequenced and directly joined by ligation to the human constant light (Ig κ) and heavy (IgG1) chain cDNAs, respectively, using unique restriction sites introduced at the variable and constant region junctions without altering the original amino acid residue sequence (Umana, P., et al., Nat Biotechnol. 17:176-180 (1999); Reff, M. E., et al., Blood 83:435-445 (1994)). Each full-length cDNA was separately subcloned into pcDNA3.1(+) (Invitrogen, Leek, The Netherlands) yielding mammalian expression vectors for chimeric C2B8 light (pC2B8L) and heavy (pC2B8H) chains.
- Production of IDEC-C2B8 in CHO cells expressing different levels of GnTIII. Establishment of two CHO cell lines, CHO-tet-GnTIII expressing different levels of GnTIII depending on the tetracycline concentration in the culture medium; and CHO-tTA, the parental cell line that does not express GnTIII has been described previously (Umana, P., et al., Nat Biotechnol. 17:176-180 (1999); Umana, P., et al., Biotechnol Bioeng. 65:542-549 (1999)). Each cell line was cotranfected with vectors pC2B8L, pC2B8H, and pZeoSV2(+) (for Zeocin resistance; Invitrogen, Leek, The Netherlands) using a calcium phosphate method. Zeocin resistant clones were transferred to a 96-well plate and assayed for IDEC-C2B8 production using an ELISA assay specific for the human constant region (4). Three IDEC-C2B8 samples were obtained from parallel cultures of a selected clone (CHO-tet-GnTIII-C2B8), differing only in the tetracycline concentration added to the medium (25, 50 and 2000 ng/mL respectively). Culture supernatants were harvested in the late exponential phase. An additional antibody sample was obtained from a CHO-tTA-derived clone, CHO-tTA-C2B8, cultured under identical conditions but without adding tetracycline to the medium. Antibody samples were purified from culture medium by protein A affinity chromatography and buffer exchanged to PBS on a cation exchange column as previously described (Umana, P., et al., Nat Biotechnol. 17:176-180 (1999)). Antibody concentration was measured using a fluorescence-based kit from Molecular Probes (Leiden, The Netherlands) with Rituximab used as standard.
- Indirect immunofluorescence. CD20-positive cells (SB cells; ATCC deposit no. ATCC CCL120) and CD20-negative cells (HSB cells; ATCC deposit no. ATCC CCL120.1) were each incubated for 1 h with 2.5 μg/ml of CHO-tet-GnTIII-derived IDEC-C2B8 antibody in Hank's balanced salt solution (GibcoBRL, Basel, Switzerland) and 2% bovine serum albumin fraction V (Roche Diagnostics, Rotkreuz, Switzerland) (HBSSB). As a negative control HBSSB was used instead of C2B8 antibody. A FITC-conjugated, anti-human Fc polyclonal antibody was used as a secondary antibody (SIGMA, St. Louis) for all samples. Cells were examined using a Leica fluorescence microscope (Wetzlar, Germany).
- Oligosaccharide profiling by MALDI/TOF-MS. Neutral, N-linked oligosaccharides were derived from C2B8 antibody samples, MabThera™ (European counterpart of Rituximab; kind gift from R. Stahel, Universit{dot over (a)}tspital, Switzerland), C2B8-25t, C2B8-50t, C2B8-2000t, and C2B8-nt, (100 μg each) as previously described (Umana, P., et al., Nat Biotechnol. 17:176-180 (1999)). Briefly, the antibody samples were first treated with Arthrobacter ureafaciens sialidase (Oxford Glycosciences, Abingson, UK) to remove any sialic acid monosaccharide residues. Neutral N-linked oligosaccharides were then released from the desialylated antibody samples using peptide-N-glycosidase F (Oxford Glycosciences), purified using micro-columns, and analyzed by MALDI/TOF-MS in an Elite Voyager 400 spectrometer (Perseptive Biosystems, Farmingham, Mass.).
- ADCC Activity Assay. Peripheral blood mononuclear cells (PBMC) were separated from heparinated fresh human blood (in all experiments obtained from the same healthy donor) by centrifugation over a Ficoll-Paque (Pharmacia Biotech, Dübendorf, Switzerland) gradient. PBMC (effector) were depleted of monocytes by plastic adherence. CD20-positive SB (target) cells, were labeled for 90 min with 100 μCi 51Cr (Amersham, Dubendorf, Switzerland) at 37° C., washed twice in RPMI (GibcoBRL, Basel, Switzerland) and resuspended at a concentration of 105 cells/ml. Fifty microliters of C2B8 mAb diluted in RPMI medium was added to 100 μl SB cells (10,000 cells/well) in a 96-well round bottom microtiter plate (Greiner, Langenthal, Switzerland), centrifuged at 50×g for 1 min, and incubated for 1 h at 4° C. Subsequently, 50 μl of effector cell (suspended at 2×107 cells/ml in RPMI medium) were added to each 96-well yielding a final E:T ratio of 100. Plates were incubated for 4 h at 37° C. and 5% CO2, supernatant was harvested with a Skatron harvesting system (Skatron Instruments, Sterling, Va.) and counted (ER, experimental release) in a Cobra 05005 γ counter (Canberra Packard, Meriden, Conn.). Maximum (MR) and spontaneous (SR) releases were obtained by adding, instead of C2B8 mAb, 100 μl of 1% Nonidet (Sigma, St. Louis) or 100 μl of RPMI medium, respectively, to 100 μl labeled target cells. All data points were performed in triplicate. Specific lysis (%) was calculated with the following formula: (ER−SR)/(MR−SR)×100.
- Results and Discussion
- Production of IDEC-C2B8 and verification of specific antigen binding. CHO-tet-GnTIII cells, with stable, tetracycline-regulated expression of GnTIII and stable, constitutive expression of IDEC-C2B8, were established and scaled-up for production of a set of antibody samples. During scale-up, parallel cultures from the same clone were grown under three different tetracycline concentrations, 25, 50 and 2000 ng/ml. These levels of tetracycline had previously been shown to result in different levels of GnTIII and bisected oligosaccharides (Umana, P., et al., Nat Biotechnol. 17:176-180(1999); Umana, P., et al., Biotechnol Bioeng. 65:542-549 (1999)). A C2B8-producing, control cell line that does not express GnTIII was also established and cultured under the same conditions as for the three parallel cultures of CHO-tet-GnTIII. After Protein A-affinity chromatography, mAb purity was estimated to be higher than 95% by SDS-PAGE and Coomassie-blue staining. The samples were named according to the tetracycline concentration added to the culture medium for their production: C2B8-25t, C2B 8-50t, C2B8-2000t and C2B8-nt (i.e., no tetracycline for the non-bisected control). Sample C2B 8-25t showed specific antigen binding by indirect immunofluorescence using CD20-positive and CD20-negative cells (
FIG. 1 ), indicating that the synthesized VL and VH gene fragments were functionally correct. - Oligosaccharide profiling with MALDI/TOF-MS. The glycosylation profile of each antibody sample was analyzed by MALDI/TOF-MS of the released, neutral oligosaccharide mix. In this technique, oligosaccharides of different mass appear as separate peaks in the spectrum and their proportions are quantitatively reflected by the relative peak heights (Harvey, D. J., Rapid Common Mass Spectrom. 7:614-619 (1993); Harvey, D. J., et al., Glycoconj J. 15:333-338 (1998)). Oligosaccharide structures were assigned to different peaks based on their expected molecular masses, previous structural data for oligosaccharides derived from IgG1 mAbs produced in the same host, and information on the N-linked oligosaccharide biosynthetic pathway.
- A clear correlation was found between GnTIII expression levels (i.e., tetracycline concentration) and the amount of bisected oligosaccharides derived from the different antibody samples. As expected, MabThera™ and C2B8-nt, which are derived from hosts that do not express GnTIII, did not carry bisected oligosaccharides (
FIGS. 2A and 2B ). In contrast, bisected structures amounted up to approximately 35% of the oligosaccharides pool in sample C2B8-2000t, i.e, at a basal level of GnTIII expression. In this case, the main bisected oligosaccharide peaks were of complex type, unequivocally assigned to peaks at m/z 1689 and m/z 1851 (FIG. 2C ). The next higher GnTIII expression level, sample C2B8-50t, led to an increase in these peaks (including their associated potassium aducts at m/z 1705 and 1861) of around 20%. This increase was accompanied by a concomitant reduction of their non-bisected counterparts at m/z FIG. 2D ). At the highest GnTIII expression level, sample C2B8-25t, the main substrate for GnTIII, m/z 1486, decreased to almost base-line level, while complex bisected structures (m/z 1689 and 1851) decreased in favor of increases in peaks at m/z FIG. 2E ). These peaks can be assigned either to bisected hybrid compounds, to galactosylated complex oligosaccharides, or to a mixture of both. Their relative increase, however, is consistent with the accumulation of bisected hybrid compounds, as GnTIII overexpression can divert the biosynthetic flux at early stages of the pathway (seeFIGS. 3A and 3B ). The amount of bisected oligosaccharide structures (complex and hybrid type) reached approximately 80% for this sample. - ADCC activity of IDEC-C2B8 glycosylated variants. Different C2B8 mAb glycosylation variants were compared for ADCC activity, measured as in vitro lysis of CD20-positive SB cells. An additional mAb sample, C2B8-nt, derived from the parental cell line lacking GnTIII, was also studied. Sample C2B8-2000t produced at the basal GnTIII expression level and carrying low levels of bisected oligosaccharides was slightly more active than C2B8-nt (
FIG. 4A ). At the next higher level of GnTIII-expression, sample C2B8-50t carried approximately equal levels of bisected and non-bisected oligosaccharides, but did not mediate significantly higher target-cell lysis. However, at the lowest tetracycline concentration, sample C2B8-25t, which contained up to 80% of bisected oligosaccharide structures, was significantly more active than the rest of the samples in the whole antibody concentration range. It reached the maximal level of ADCC activity of sample C2B8-nt at a 10-fold lower antibody concentration (FIG. 4A ). Sample C2B8-25t also showed a significant increase in the maximal ADCC activity with respect to the control (50% vs. 30% lysis). - Samples C2B8-50t and C2B8-25t, bearing the highest proportions of bisected oligosaccharides, were further compared in ADCC activity to Mabthera™, the version of Rituxan™ currently marketed in Europe (
FIG. 4B ). Sample C2B8-50t showed a slight increase in activity whereas sample C2B8-25t clearly outperformed Mabthera™ at all antibody concentrations. Approximately a five to ten-fold lower concentration of C2B8-25t was required to reach the maximal ADCC activity of Mabthera™, and the maximal activity of C2B8-25t was about 25% higher than that of Mabthera™. - These results show that, in general, the in vitro ADCC activity of the C2B8 antibody correlates with the proportion of molecules carrying bisected oligosaccharides in the Fc region. We had previously reported that in the case of chCE7, an antibody with a low baseline level of ADCC activity, significant increases of activity could be obtained by increasing the fraction of bisected oligosaccharides above the levels found in naturally-occurring antibodies (Umana, P., et al., Nat Biotechnol. 17:176-180 (1999)). The same is true for the C2B8 mAb, which already has high ADCC activity in the absence of bisected oligosaccharides. In the case of chCE7, however, very large increases of ADCC activity were observed at a level of GnTIII expression where bisected oligosaccharides were predominantly of complex type (Umana, P., et al., Nat Biotechnol. 17:176-180 (1999)). For the potent C2B8 mAb, such a large boost in activity was only observed at the highest levels of GnTIII expression studied, where bisected oligosaccharides had shifted mainly to the hybrid type (
FIG. 2 ). For both mAbs, the samples with the highest activities had considerably higher levels of bisected than non-bisected oligosaccharides. Together, these observations indicate that probably both complex and hybrid bisected oligosaccharides are important for ADCC activity. - In both complex and hybrid oligosaccharides, a bisecting GlcNAc leads to a large change in oligosaccharide conformations (Balaji, P. V., et al., Int. J. Biol. Macromol. 18:101-114 (1996)). The change occurs in a part of the oligosaccharide that interacts extensively with the polypeptide in the CH2 domain (Jefferis, R., et al., Immunol Rev. 163:59-76 (1998)). Since the polypetide is relatively flexible at this location (Jefferis, R., et al., Immunol Rev. 163:59-76 (1998)), it is possible that the bisecting N-acetylglucosamine is mediating its biological effects through a conformational change in the Fc region. The potentially altered conformations would already exist in nature, as all serum IgGs carry bisected oligosaccharides. The main difference between the engineered and natural antibodies would be the proportion of molecules displaying the more active conformations.
- Various approaches for increasing the activity of unconjugated mAbs are currently under clinical evaluation, including radio-immunotherapy, antibody-dependent enzyme/prodrugtherapy, immunotoxins and adjuvant therapy with cytokines (Hjelm Skog, A., et al., Cancer Immunol Immunother. 48:463-470 (1999); Blakey, D. C., et al., Cell Biophys. 25: 175-183 (1994); Wiseman, G. A., et al., Clin Cancer Res. 5:3281s-3296s (1999); Hank, J. A., et al., Cancer Res. 50:5234-5239 (1990)). These technologies can give large increases in activity, but they can also lead to significantly higher side effects, elevated production costs and complex logistics from production to administration to patients when compared to unconjugated mAbs. The technology presented here offers an alternative way to obtain increases in potency while maintaining a simple production process, and should be applicable to many unconjugated mAbs.
- 1. Cell Culture
- SP2/0 mouse myeloma cells producing chG250 chimeric mAb (wt-chG250-SP2/0 cells) were grown in standard cell culture medium supplemented with 1:100 (v/v) penicillin/streptomycin/antimycotic solution (SIGMA, Buchs, Switzerland). Cells were cultured at 37° C. in a 5% CO2 humidified atmosphere in Tissue Culture Flasks. Medium was changed each 3-4 days. Cells were frozen in culture medium containing 10% DMSO.
- 2. Generation of SP2/0 Cells with pGnTIII-puro Expression
- wt-chG250-SP2/O myeloma cells were transfected by electroporation with a vector for constitutive expression of GnTIII operatively linked via an IRES to a puromycin resistance gene. 24 hours before electroporation culture medium was changed and cells were seeded at 5×105 cells/ml. Seven million cells were centrifuged for 4 min at 1300 rpm at 4° C. Cells were washed with 3 mL new medium and centrifuged again. Cells were resuspended in a volume of 0.3-0.5 ml of reaction mix, containing 1.25% (v/v) DMSO and 20-30 μg DNA in culture medium. The electroporation mix was then transferred to a 0.4 cm cuvette and pulsed at low voltage (250-300 V) and high capacitance (960 μF) using Gene Pulser from Bio Rad. After electroporation cells were quickly transferred to 6 mL 1.25% (v/v) DMSO culture medium in a T25 culture flask and incubated at 37° C. Stable integrants were selected by applying 2 μg/mL puromycin to the medium two days after electroporation. After 2-3 weeks a stable, puromycin-resitant mixed population was obtained. Single-cell derived clones were obtained via FACS and were subsequently expanded and maintained under puromycin selection.
- 3. Western Blot
- Puromycin-resistant clones were screened for GnTIII expression by Western blotting. The Western blots clearly showed that clones 5H12, 4E6 and 4E8 were expressing the highest levels of GnTIII. 5G2 also showed a GnTIII band of middle intensity, whereas 2F1, 3D3 and 4G3 had the lowest band intensities, therefore expressing lower amounts of GnTIII (
FIG. 5 ). - 4. Production and Purification of chG250 Monoclonal Antibody from Seven GnTIII-Expressing Clones Including Wild Type
- Clones 2F1, 3D3, 4E6, 4E8, 4G3, 5G2, 5H12 and the wild type (wt-chG250-SP2/0 cells) were seeded at 3×105 cells/mL in a total volume of 130 ml culture medium, and cultivated in single Triple-flasks. Cells used for seeding were all in full exponential growth phase, therefore cells were considered to be at the same growth state when the production batches started. Cells were cultivated for 4 days. Supernatants containing the antibody were collected in the late exponential growth phase to ensure reproducibility. The chG250 monoclonal antibody was purified in two chromatographic steps. Culture supernatants containing the chG250 monoclonal antibody derived from each batch were first purified using a HiTrap Protein A affinity chromatography. Protein A is highly specific for the human IgG F, region. Pooled samples from the protein A eluate were buffer exchanged to PBS by cation-exchange chromatography on a
Resource S 1 ml column (Amersham Pharmacia Biotech). Final purity was judged to be higher than 95% from SDS-staining and Coomassie blue staining (FIG. 6 ). The concentration of each sample was determined with a standard calibration curve using wild type antibody with known concentration. - 5. Oligosaccharide Profiling of mAb Preparations Derived from the Seven Clones Expressing Different GnTIII Levels
- Oligosaccharide profiles were obtained by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI/TOF-MS), which accurately provides the molecular masses of the different oligosaccharide structures. This technique allows a quantitative analysis of proportions between different oligosaccharide structures within a mixture. Neutral oligosaccharides appeared predominantly as [M+Na+] ions, however sometimes they were accompanied by-smaller [M+K+] ions, leading to an increase in mass of m/z of 16. The percentage of the structure appearing as potassium ion adducts depends on the content of the matrix and may thus vary between samples. A mixture of neutral N-linked oligosaccharides derived from each antibody preparation was analyzed using a 2,5-dehydrobenzoic acid (2,5-DHB) as matrix. Some of the peaks in the spectra were unequivocally assigned to specific oligosaccharide structures, because of known monosaccharide composition and unique mass. However, sometimes multiple structures could be assigned to a particular mass. MALDI enables the determination of the mass and cannot distinguish between isomers. Knowledge of the biosynthetic pathway and previous structural data enable, in most cases, the assignment of an oligosaccharide structure to a peak in the spectrum.
- Oligosaccharides derived from the mAb sample produced in wt-chG250-SP2/0 cell line, that does not express GnTIII, contained nonbisected biantennary complex (m/z 1486) and mono- or di-galactosylated nonbisected biantennary complex structures (
FIG. 7A ), both α(1,6)-fucosylated in the core region (peaks m/z - Expression of GnTIII generated bisected Fc-associated oligosaccharide structures of two types: complex or hybrid. Complex bisected oligosaccharides were unequivocally assigned to peaks at m/
z z z 1648, had the lowest values for the clones expressing the highest GnTIII levels (clones 4E6, 4E8, 5G2 and 5H12). These two peaks decreased in favor of the accumulation of bisected complex and bisected hybrid type oligosaccharides (FIGS. 7A-7D and 8A-8D). The percentage of bisected complex oligosaccharides was higher for the samples derived from the clones expressing lower amounts of GnTIII. This is consistent with the fact that a higher GnTIII expression level probably shifts the biosynthetic flux to bisected hybrid structures, thereby decreasing the relative proportions of complex and complex bisected compound. For bisected hybrid structures, two possible structures could sometimes be assigned to a single peak. Therefore, some assumptions were made in order to approximate the percentage of these structures over the total oligosaccharide pool. Peaks m/z FIGS. 7A-7D and 8A-8D). However to assign a specific structure to peaks m/z - 6. Measurement of Antibody Mediated Cytotoxic Activity by Calcein-AM Retention
- The Calcein-AM retention method of measuring cytotoxicity measures the dye fluorescence remaining in the cells after incubation with the antibody. Four million G250 antigen-positive cells (target) were labelled with 10 μM Calcein-AM (Molecular Probes, Eugene, Oreg.) in 1.8 mL RPMI-1640 cell culture medium (GIBCO BRL, Basel, Switzerland) supplemented with 10% fetal calf serum for 30 min at 37° C. in a 5% CO2 humidified atmosphere. The cells were washed twice in culture medium and resuspended in 12 mL AIMV serum free medium (GIBCO BRL, Basel, Switzerland). Labelled cells were then transferred to U-bottom 96-wells (30,000 cells/well) and incubated in triplicate with different concentrations of antibody for 1 hour at 4° C. Peripheral blood mononuclear cells (PBMC) were separated from heparinated fresh human blood (in all experiments obtained from the same healthy donor) by centrifugation over a Ficoll-Paque (Pharmacia Biotech, Dübendorf, Switzerland) gradient. PBMCs were added in triplicate wells in a 50 μL volume, yielding an effector to target ratio (E:T ratio) of 25:1 and a final volume of 200 μL. The 96-well plate was then incubated for 4 hours at 37° C. in a 5% CO2 atmosphere. Thereafter the 96-well plate was centrifuged at 700×g for 5 min and the supernatants were discarded. The cell pellets were washed twice with Hank's balanced salt solution (HBSS) and lysed in 200 μL 0.05M sodium borate, pH 9, 0.1% Triton X-100. Retention of the fluorescent dye in the target cells was measured with a FLUOstar microplate reader (BMG LabTechnologies, Offenburg, Germany). The specific lysis was calculated relative to a total lysis control, resulting from exposure of the target cells to saponin (200 mg/mL in AIMV; SIGMA, Buchs, Switzerland) instead of exposure to antibody. Specific lysis (%) was calculated with the following formula:
where Fmed represents the fluorescence of target cells treated with medium alone and considers unspecific lysis by PMBCs, Fexp represents the fluorescence of cells treated with antibody and Fdet represents the fluorescence of cells treated with saponin instead of antibody. - To determine the effect of modified glycosylation variants of chG250 on the in vitro ADCC activity, G250 antigen-positive target cells were cultured with PBMCs with and without chG250 antibody samples at different concentrations. The cytotoxicity of unmodified chG250 antibody derived from the wild type cell line was compared with two antibody preparations derived from two cell lines (3D3, 5H12) expressing intermediate and high GnTIII levels, respectively (see
FIG. 5 ). - Unmodified chG250 antibody did not mediate significant ADCC activity over the entire concentration range used in the assay (the activity was not significantly different from background). Augmented ADCC activity (close to 20%, see
FIG. 9 ) at 2 μg/mL was observed with the antibody sample derived from clone 3D3, which expressed intermediate GnTIII levels. The cytotoxic activity of this antibody samples did not grow at higher antibody concentrations. As expected the antibody preparation derived from clone 5H12 showed a striking increase over samples 3D3 and unmodified antibody in its ability to mediate ADCC against target cells. The maximal ADCC activity of this antibody preparation was around 50% and was remarkable in mediating significant ADCC activity at 125-fold less concentrated when comparing with the unmodified control sample. - Autoimmune thrombocytopenia in chronic graft-versus-host disease represents an instance of B-cell dysregulation leading to clinical disease. To treat immune-mediated thrombocytopenia in a subject with chronic graft-versus-host disease, an anti-CD20 chimeric monoclonal antibody prepared by the methods of the present invention and having increased ADCC is administered to the subject as described in Ratanatharathorn, V. et al., Ann. Intern. Med. 133(4):275-79 (2000) (the entire contents of which is hereby incorporated by reference). Specifically, a weekly infusion of the antibody, 375 mg/m2 is administered to the subject for 4 weeks. The antibody therapy produces a marked depletion of B cells in the peripheral blood and decreased levels of platelet-associated antibody.
- Immune-mediated, acquired pure red cell aplasia (PRCA) is a rare disorder frequently associated with other autoimmune phenomena To treat immune-mediated, acquired pure red cell aplasia in a subject, an anti-CD20 chimeric monoclonal antibody prepared by the methods of the present invention and having increased ADCC is administered to the subject as described in Zecca, M. et al., Blood 12:3995-97 (1997) (the entire contents of which are hereby incorporated by reference). Specifically, a subject with PRCA and autoimmune hemolytic anemia is given two doses of antibody, 375 mg/m2, per week. After antibody therapy, substitutive treatment with intravenous immunoglobulin is initiated. This treatment produces a marked depletion of B cells and a significant rise in reticulocyte count accompanied by increased hemoglobin levels.
- It will be clear that the invention may be practiced otherwise than as particularly described in the foregoing description and examples. Numerous modifications and variations of the present invention are possible in light of the above teachings and, therefore, are within the scope of the appended claims.
- The entire disclosure of all publications (including patents, patent applications, journal articles, laboratory manuals, books, or other documents) cited herein are hereby incorporated by reference.
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Priority Applications (4)
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US13/196,724 US8999324B2 (en) | 1998-04-20 | 2011-08-02 | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
US14/665,191 US9321843B2 (en) | 1998-04-20 | 2015-03-23 | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
US15/080,020 US9631023B2 (en) | 1998-04-20 | 2016-03-24 | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
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US11/199,232 US8021856B2 (en) | 1998-04-20 | 2005-08-09 | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
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US11/199,232 Continuation US8021856B2 (en) | 1998-04-20 | 2005-08-09 | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
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US13/196,724 Division US8999324B2 (en) | 1998-04-20 | 2011-08-02 | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
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US15/080,020 Expired - Fee Related US9631023B2 (en) | 1998-04-20 | 2016-03-24 | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050123546A1 (en) * | 2003-11-05 | 2005-06-09 | Glycart Biotechnology Ag | Antigen binding molecules with increased Fc receptor binding affinity and effector function |
US20090060911A1 (en) * | 2000-04-13 | 2009-03-05 | The Rockefeller University | Enhancement of antibody-mediated immune responses |
US20100081195A1 (en) * | 2008-09-26 | 2010-04-01 | Eureka Therapeutics, Inc. | Modified Host Cells and Uses Thereof |
Families Citing this family (915)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6136311A (en) | 1996-05-06 | 2000-10-24 | Cornell Research Foundation, Inc. | Treatment and diagnosis of cancer |
ES2434961T5 (en) * | 1998-04-20 | 2018-01-18 | Roche Glycart Ag | Antibody glycosylation engineering to improve antibody-dependent cell cytotoxicity |
ATE475714T1 (en) | 1998-12-09 | 2010-08-15 | Phyton Holdings Llc | METHOD FOR PRODUCING GLYCOPROTEIN WITH HUMAN GLYCOSYLATION IMAGE |
ATE464907T1 (en) | 1999-02-17 | 2010-05-15 | Csl Ltd | IMMUNOGENIC COMPLEXES AND METHODS RELATING THEM |
ATE365219T1 (en) | 1999-10-26 | 2007-07-15 | Plant Res Int Bv | MAMMAL-TYPE GLYCOLIZATION IN PLANTS |
US7598055B2 (en) | 2000-06-28 | 2009-10-06 | Glycofi, Inc. | N-acetylglucosaminyltransferase III expression in lower eukaryotes |
US7449308B2 (en) | 2000-06-28 | 2008-11-11 | Glycofi, Inc. | Combinatorial DNA library for producing modified N-glycans in lower eukaryotes |
US8697394B2 (en) * | 2000-06-28 | 2014-04-15 | Glycofi, Inc. | Production of modified glycoproteins having multiple antennary structures |
ATE309385T1 (en) * | 2000-06-28 | 2005-11-15 | Glycofi Inc | METHOD FOR PRODUCING MODIFIED GLYCOPROTEINS |
EP1356068B1 (en) | 2001-01-19 | 2014-10-29 | Phyton Holdings, LLC | Method for secretory production of glycoprotein having human-type sugar chain using plant cell |
JP2005500018A (en) * | 2001-04-02 | 2005-01-06 | アイデック ファーマスーティカルズ コーポレイション | Recombinant antibody coexpressed with GnTIII |
KR20100018071A (en) | 2001-08-03 | 2010-02-16 | 글리카트 바이오테크놀로지 아게 | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
WO2003055514A1 (en) * | 2001-12-21 | 2003-07-10 | Antigenics Inc. | Compositions comprising immunoreactive reagents and saponins, and methods of use thereof |
US20070148171A1 (en) * | 2002-09-27 | 2007-06-28 | Xencor, Inc. | Optimized anti-CD30 antibodies |
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 |
US7317091B2 (en) | 2002-03-01 | 2008-01-08 | Xencor, Inc. | Optimized Fc variants |
US20080260731A1 (en) * | 2002-03-01 | 2008-10-23 | Bernett Matthew J | Optimized antibodies that target cd19 |
US8188231B2 (en) | 2002-09-27 | 2012-05-29 | Xencor, Inc. | Optimized FC variants |
US20040132101A1 (en) | 2002-09-27 | 2004-07-08 | Xencor | Optimized Fc variants and methods for their generation |
AU2003219402B2 (en) | 2002-03-19 | 2008-05-08 | Stichting Dienst Landbouwkundig Onderzoek | GnTIII (UDP-n-acethylglucosamine:beta-D mannoside beta (1,4)-N-acethylglucosaminy ltransferase III) expression in plants |
KR101196023B1 (en) | 2002-03-19 | 2012-10-30 | 스티칭 디엔스트 랜드보위쿤디그 온데조에크 | Optimizing Glycan Processing in Plants |
US20060235208A1 (en) * | 2002-09-27 | 2006-10-19 | Xencor, Inc. | Fc variants with optimized properties |
CL2003002461A1 (en) | 2002-11-27 | 2005-01-07 | Dow Chemical Company Agroscien | IMMUNOGLOBULIN THAT UNDERSTANDS AT LEAST ONE AFUCOSILATED GLICAN, COMPOSITION THAT CONTAINS IT, NUCLEOTIDIC SEQUENCE AND VECTOR THAT UNDERSTANDS IT, PROCEDURE TO PRODUCE IMMUNOGLOBULIN SAID IN PLANTS. |
US7638270B2 (en) * | 2003-01-24 | 2009-12-29 | Agensys, Inc. | Nucleic acids and corresponding proteins entitled 254P1D6B useful in treatment and detection of cancer |
US7332299B2 (en) | 2003-02-20 | 2008-02-19 | Glycofi, Inc. | Endomannosidases in the modification of glycoproteins in eukaryotes |
US8388955B2 (en) * | 2003-03-03 | 2013-03-05 | Xencor, Inc. | Fc variants |
US20070275460A1 (en) * | 2003-03-03 | 2007-11-29 | Xencor.Inc. | Fc Variants With Optimized Fc Receptor Binding Properties |
US20090010920A1 (en) | 2003-03-03 | 2009-01-08 | Xencor, Inc. | Fc Variants Having Decreased Affinity for FcyRIIb |
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 |
AR044388A1 (en) | 2003-05-20 | 2005-09-07 | Applied Molecular Evolution | CD20 UNION MOLECULES |
US9714282B2 (en) | 2003-09-26 | 2017-07-25 | Xencor, Inc. | Optimized Fc variants and methods for their generation |
US8101720B2 (en) | 2004-10-21 | 2012-01-24 | Xencor, Inc. | Immunoglobulin insertions, deletions and substitutions |
US20050249723A1 (en) * | 2003-12-22 | 2005-11-10 | Xencor, Inc. | Fc polypeptides with novel Fc ligand binding sites |
EP2168986A3 (en) | 2004-02-19 | 2010-07-28 | Genentech, Inc. | CDR-repaired antibodies |
AR048098A1 (en) * | 2004-03-15 | 2006-03-29 | Wyeth Corp | CALIQUEAMYCIN CONJUGATES |
EP2053062A1 (en) * | 2004-03-24 | 2009-04-29 | Xencor, Inc. | Immunoglobin variants outside the Fc region |
AU2005247303A1 (en) * | 2004-04-16 | 2005-12-08 | Genentech, Inc. | Treatment of polychondritis and mononeuritis multiplex with anti-CD20 antibodies |
RU2384345C2 (en) | 2004-06-04 | 2010-03-20 | Дженентек, Инк. | Method of treating multiple sclerosis |
NZ545720A (en) * | 2004-07-09 | 2010-01-29 | Chugai Pharmaceutical Co Ltd | Anti-glypican 3 antibody |
CA2573505C (en) * | 2004-07-14 | 2013-06-25 | Igeneon Krebs-Immuntherapie Forschungs-Und Entwicklungs-Ag | N-glycosylated antibody |
US20150010550A1 (en) | 2004-07-15 | 2015-01-08 | Xencor, Inc. | OPTIMIZED Fc VARIANTS |
US20060074225A1 (en) * | 2004-09-14 | 2006-04-06 | Xencor, Inc. | Monomeric immunoglobulin Fc domains |
CN101087807A (en) | 2004-10-05 | 2007-12-12 | 健泰科生物技术公司 | Method for treating vasculitis |
JO3000B1 (en) | 2004-10-20 | 2016-09-05 | Genentech Inc | Antibody Formulations. |
WO2006046751A1 (en) * | 2004-10-26 | 2006-05-04 | Chugai Seiyaku Kabushiki Kaisha | Anti-glypican 3 antibody having modified sugar chain |
US8367805B2 (en) | 2004-11-12 | 2013-02-05 | Xencor, Inc. | Fc variants with altered binding to FcRn |
EP2314618A3 (en) | 2004-11-12 | 2011-10-19 | 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 |
US8546543B2 (en) | 2004-11-12 | 2013-10-01 | Xencor, Inc. | Fc variants that extend antibody half-life |
CA2595169A1 (en) * | 2005-01-12 | 2006-07-20 | Xencor, Inc. | Antibodies and fc fusion proteins with altered immunogenicity |
KR20190110637A (en) | 2005-01-21 | 2019-09-30 | 제넨테크, 인크. | Fixed dosing of her antibodies |
PL1871805T3 (en) | 2005-02-07 | 2020-03-31 | Roche Glycart Ag | Antigen binding molecules that bind egfr, vectors encoding same, and uses thereof |
SG170006A1 (en) * | 2005-02-18 | 2011-04-29 | Medarex Inc | Monoclonal antibodies against cd30 lacking fucosyl residues |
SI1850874T1 (en) | 2005-02-23 | 2014-01-31 | Genentech, Inc. | Extending time to disease progression or survival in ovarian cancer patients using pertuzumab |
CA2601858A1 (en) * | 2005-03-25 | 2006-09-28 | Glycart Biotechnology Ag | Antigen binding molecules directed to mcsp and having increased fc receptor binding affinity and effector function |
JO3058B1 (en) | 2005-04-29 | 2017-03-15 | Applied Molecular Evolution Inc | Anti-IL-6 Antibodies,Compositions,Methods and uses |
CA2605781A1 (en) * | 2005-05-09 | 2007-04-12 | Glycart Biotechnology Ag | Antigen binding molecules having modified fc regions and altered binding to fc receptors |
CN101282993A (en) * | 2005-06-02 | 2008-10-08 | 阿斯利康公司 | Antibodies directed to cd20 and uses thereof |
WO2006133148A2 (en) * | 2005-06-03 | 2006-12-14 | Genentech, Inc. | Method of producing antibodies with modified fucosylation level |
EP1896073B1 (en) | 2005-06-30 | 2013-03-06 | Janssen Biotech, Inc. | Anti-il-23 antibodies, compositions, methods and uses |
US9580506B2 (en) | 2005-07-21 | 2017-02-28 | Genmab A/S | Potency assays for antibody drug substance binding to an Fc receptor |
WO2007024743A2 (en) * | 2005-08-19 | 2007-03-01 | Centocor, Inc. | Proteolysis resistant antibody preparations |
NO345919B1 (en) * | 2005-08-26 | 2021-10-18 | Roche Glycart Ag | Modified antigen-binding molecules with altered cell signaling activity |
CA2624189A1 (en) * | 2005-10-03 | 2007-04-12 | Xencor, Inc. | Fc variants with optimized fc receptor binding properties |
CA2625998C (en) | 2005-10-06 | 2015-12-01 | Xencor, Inc. | Optimized anti-cd30 antibodies |
US20070087005A1 (en) * | 2005-10-14 | 2007-04-19 | Lazar Gregory A | Anti-glypican-3 antibody |
MY149159A (en) | 2005-11-15 | 2013-07-31 | Hoffmann La Roche | Method for treating joint damage |
JP2009518314A (en) | 2005-12-02 | 2009-05-07 | ジェネンテック・インコーポレーテッド | Compositions and methods for the treatment of diseases and disorders associated with cytokine signaling comprising antibodies that bind IL-22 and antibodies that bind IL-22R |
RS53685B1 (en) | 2005-12-29 | 2015-04-30 | Janssen Biotech Inc. | Human anti-il-23 antibodies, compositions, methods and uses |
AU2007248444B2 (en) | 2006-01-05 | 2012-10-25 | Genentech, Inc. | Anti-EphB4 antibodies and methods using same |
BRPI0707290A2 (en) | 2006-01-17 | 2011-08-16 | Biolex Therapeutics Inc | compositions and methods for humanization and optimization of n-glycans in plants |
US20070166306A1 (en) * | 2006-01-17 | 2007-07-19 | Fey Georg H M | Anti-CD19 antibody composition and method |
DK1991273T4 (en) * | 2006-02-10 | 2022-02-07 | Life Technologies Corp | LABELING AND DETECTION OF POST-TRANSLATIONALLY MODIFIED PROTEINS |
AR059851A1 (en) | 2006-03-16 | 2008-04-30 | Genentech Inc | ANTIBODIES OF EGFL7 AND METHODS OF USE |
JP2009531324A (en) | 2006-03-20 | 2009-09-03 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | Engineered anti-prostatic stem cell antigen (PSCA) antibody for cancer targeting |
RU2436796C9 (en) | 2006-05-30 | 2013-12-27 | Дженентек, Инк. | Antibodies and immunoconjugates and their application |
EP2032604A2 (en) | 2006-06-06 | 2009-03-11 | Genentech, Inc. | Anti-dll4 antibodies and methods using same |
PL2468770T3 (en) | 2006-07-14 | 2018-07-31 | Ac Immune S.A. | Humanized antibody against amyloid beta. |
RS53160B (en) | 2006-07-14 | 2014-06-30 | Ac Immune S.A. | Humanized antibody against amyloid beta |
EP2046809B1 (en) | 2006-07-19 | 2016-12-07 | The Trustees Of The University Of Pennsylvania | Wsx-1/il-27 as a target for anti-inflammatory responses |
AR062223A1 (en) * | 2006-08-09 | 2008-10-22 | Glycart Biotechnology Ag | MOLECULES OF ADHESION TO THE ANTIGEN THAT ADHER TO EGFR, VECTORS THAT CODE THEM, AND THEIR USES OF THESE |
DK2383297T5 (en) | 2006-08-14 | 2022-07-04 | Xencor Inc | Optimized antibodies directed against CD19 |
US8911964B2 (en) | 2006-09-13 | 2014-12-16 | Abbvie Inc. | Fed-batch method of making human anti-TNF-alpha antibody |
CA2910619A1 (en) | 2006-09-13 | 2008-03-20 | Abbvie Inc. | Cell culture improvements |
CA2660795C (en) * | 2006-09-18 | 2014-11-18 | Xencor, Inc. | Optimized antibodies that target hm1.24 |
JP5298021B2 (en) * | 2006-10-12 | 2013-09-25 | ジェネンテック, インコーポレイテッド | Antibodies against lymphotoxin-α |
KR101541550B1 (en) | 2006-10-27 | 2015-08-04 | 제넨테크, 인크. | Antibodies and immunoconjugates and uses therefor |
NZ578064A (en) * | 2006-12-01 | 2012-01-12 | Medarex Inc | Human antibodies that bind cd22 and uses thereof |
RU2009133784A (en) | 2007-02-09 | 2011-03-20 | Дженентек, Инк. (Us) | ANTI-Robo4-ANTIBODIES AND THEIR APPLICATIONS |
CN103432580A (en) | 2007-03-02 | 2013-12-11 | 健泰科生物技术公司 | Predicting response to a HER dimerisation inhibitor based on low HER3 expression |
US20100028951A1 (en) * | 2007-03-07 | 2010-02-04 | Stephen Hamilton | Production of glycoproteins with modified fucosylation |
US7960139B2 (en) | 2007-03-23 | 2011-06-14 | Academia Sinica | Alkynyl sugar analogs for the labeling and visualization of glycoconjugates in cells |
NZ593203A (en) | 2007-04-17 | 2012-08-31 | Stichting Dienst Landbouwkundi | Mammalian-type glycosylation in plants by expression of non-mammalian glycosyltransferases by inserting a polypeptide coding for a non-mammalian beta1,4-galactosyltransferase |
PL2068927T3 (en) | 2007-05-14 | 2016-06-30 | Medimmune Llc | Methods of reducing eosinophil levels |
SI2171090T1 (en) | 2007-06-08 | 2013-07-31 | Genentech, Inc. | Gene expression markers of tumor resistance to her2 inhibitor treatment |
SG182192A1 (en) | 2007-06-12 | 2012-07-30 | Ac Immune Sa | Humanized antibodies to amyloid beta |
HUE031698T2 (en) * | 2007-06-15 | 2017-07-28 | Medicago Inc | Modifying glycoprotein production in plants |
US7580304B2 (en) * | 2007-06-15 | 2009-08-25 | United Memories, Inc. | Multiple bus charge sharing |
EP2188302B1 (en) | 2007-07-09 | 2017-11-01 | Genentech, Inc. | Prevention of disulfide bond reduction during recombinant production of polypeptides |
WO2009032949A2 (en) | 2007-09-04 | 2009-03-12 | The Regents Of The University Of California | High affinity anti-prostate stem cell antigen (psca) antibodies for cancer targeting and detection |
EP3415529B1 (en) | 2007-09-26 | 2020-11-04 | Chugai Seiyaku Kabushiki Kaisha | Modified antibody constant region |
US20100297012A1 (en) | 2007-10-05 | 2010-11-25 | Andrea Pfeifer | Humanized antibody |
AU2008323770B2 (en) | 2007-11-07 | 2014-08-07 | Genentech, Inc. | Compositions and methods for treatment of microbial disorders |
TWI580694B (en) | 2007-11-30 | 2017-05-01 | 建南德克公司 | Anti-vegf antibodies |
WO2009086072A2 (en) * | 2007-12-21 | 2009-07-09 | Genentech, Inc. | Therapy of rituximab-refractory rheumatoid arthritis patients |
ES2742268T3 (en) | 2007-12-26 | 2020-02-13 | Xencor Inc | Fc variants with altered FcRn binding |
CN101945892B (en) * | 2007-12-26 | 2017-11-24 | 生物测试股份公司 | For the method and reagent of the targeting for improving the tumour cell to expressing CD138 |
WO2009080831A1 (en) * | 2007-12-26 | 2009-07-02 | Biotest Ag | Method of decreasing cytotoxic side-effects and improving efficacy of immunoconjugates |
CA2710453C (en) * | 2007-12-26 | 2019-07-02 | Biotest Ag | Agents targeting cd138 and uses thereof |
CA2710471C (en) | 2007-12-26 | 2018-06-05 | Biotest Ag | Immunoconjugates targeting cd138 and uses thereof |
TWI472339B (en) | 2008-01-30 | 2015-02-11 | Genentech Inc | Composition comprising antibody that binds to domain ii of her2 and acidic variants thereof |
WO2009134738A1 (en) * | 2008-04-29 | 2009-11-05 | Genentech, Inc. | Responses to immunizations in rheumatoid arthritis patients treated with a cd20 antibody |
KR101054362B1 (en) * | 2008-07-03 | 2011-08-05 | 재단법인 목암생명공학연구소 | How to reduce the fucose content of recombinant protein |
US8680020B2 (en) | 2008-07-15 | 2014-03-25 | Academia Sinica | Glycan arrays on PTFE-like aluminum coated glass slides and related methods |
TW201014605A (en) | 2008-09-16 | 2010-04-16 | Genentech Inc | Methods for treating progressive multiple sclerosis |
CN102245208B (en) | 2008-10-14 | 2016-03-16 | 霍夫曼-拉罗奇有限公司 | immunoglobulin variants and uses thereof |
TW201024318A (en) | 2008-10-20 | 2010-07-01 | Abbott Lab | Isolation and purification of antibodies using protein A affinity chromatography |
CA2738499A1 (en) | 2008-10-20 | 2010-04-29 | Abbott Laboratories | Viral inactivation during purification of antibodies |
CA2744158A1 (en) | 2008-11-22 | 2010-05-27 | Genentech, Inc. | Anti-angiogenesis therapy for the treatment of breast cancer |
TW201029662A (en) | 2008-12-19 | 2010-08-16 | Glaxo Group Ltd | Novel antigen binding proteins |
WO2010075249A2 (en) | 2008-12-22 | 2010-07-01 | Genentech, Inc. | A method for treating rheumatoid arthritis with b-cell antagonists |
JP6039183B2 (en) | 2008-12-23 | 2016-12-07 | ジェネンテック, インコーポレイテッド | Immunoglobulin variants with altered binding to protein A |
WO2010096486A1 (en) | 2009-02-17 | 2010-08-26 | Cornell Research Foundation, Inc. | Methods and kits for diagnosis of cancer and prediction of therapeutic value |
WO2010098865A1 (en) * | 2009-02-26 | 2010-09-02 | Gt Life Sciences, Inc. | Mammalian cell line models and related methods |
MX2011009729A (en) | 2009-03-20 | 2011-10-14 | Genentech Inc | Bispecific anti-her antibodies. |
EP2679600A1 (en) | 2009-03-25 | 2014-01-01 | Genentech, Inc. | Anti-FGFR3 antibodies and methods using same |
TWI504409B (en) | 2009-03-25 | 2015-10-21 | Genentech Inc | Novel anti-α5β1 antibodies and uses thereof |
CN102573916B (en) * | 2009-05-06 | 2017-05-17 | 生物测试股份公司 | Uses of immunoconjugates targeting cd138 |
EP2435476A4 (en) | 2009-05-27 | 2013-04-17 | Synageva Biopharma Corp | Avian derived antibodies |
US9676845B2 (en) | 2009-06-16 | 2017-06-13 | Hoffmann-La Roche, Inc. | Bispecific antigen binding proteins |
US20100316639A1 (en) | 2009-06-16 | 2010-12-16 | Genentech, Inc. | Biomarkers for igf-1r inhibitor therapy |
WO2011014457A1 (en) | 2009-07-27 | 2011-02-03 | Genentech, Inc. | Combination treatments |
EP2459591B1 (en) | 2009-07-31 | 2014-08-20 | Genentech, Inc. | Inhibition of tumor metastasis using anti-g-csf-antibodies |
CN104059955A (en) | 2009-08-11 | 2014-09-24 | 弗·哈夫曼-拉罗切有限公司 | Production Of Proteins In Glutamine-free Cell Culture Media |
BR112012003346A2 (en) | 2009-08-15 | 2016-11-16 | Genentech Inc | treatment method of a previously treated metastatic breast cancer diagnostic patient previously treated metastatic breast cancer treatment kit in a human patient method to instruct a human cancer patient promotional method and commercial method |
TWI412375B (en) * | 2009-08-28 | 2013-10-21 | Roche Glycart Ag | Humanized anti-cdcp1 antibodies |
SG178567A1 (en) * | 2009-08-31 | 2012-04-27 | Roche Glycart Ag | Affinity-matured humanized anti cea monoclonal antibodies |
US9493578B2 (en) | 2009-09-02 | 2016-11-15 | Xencor, Inc. | Compositions and methods for simultaneous bivalent and monovalent co-engagement of antigens |
AU2010289400B2 (en) | 2009-09-02 | 2014-10-23 | Curis, Inc. | Mutant smoothened and methods of using the same |
CA2778481A1 (en) | 2009-10-22 | 2011-04-28 | Genentech, Inc. | Anti-hepsin antibodies and methods using same |
WO2011056497A1 (en) | 2009-10-26 | 2011-05-12 | Genentech, Inc. | Activin receptor type iib compositions and methods of use |
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 |
EP2496600A1 (en) | 2009-11-04 | 2012-09-12 | Fabrus LLC | Methods for affinity maturation-based antibody optimization |
EP2496601B1 (en) | 2009-11-05 | 2017-06-07 | F. Hoffmann-La Roche AG | Methods and composition for secretion of heterologous polypeptides |
US10087236B2 (en) | 2009-12-02 | 2018-10-02 | Academia Sinica | Methods for modifying human antibodies by glycan engineering |
US11377485B2 (en) | 2009-12-02 | 2022-07-05 | Academia Sinica | Methods for modifying human antibodies by glycan engineering |
RU2673908C2 (en) | 2009-12-02 | 2018-12-03 | Имэджинэб, Инк. | J591 minibodies and cys-diabodies for targeted delivery of human prostate specific membrane antigen (psma) and methods for their use |
TWI505836B (en) | 2009-12-11 | 2015-11-01 | Genentech Inc | Anti-vegf-c antibodies and methods using same |
MX2012007379A (en) | 2009-12-23 | 2012-08-31 | Genentech Inc | Anti-bv8 antibodies and uses thereof. |
US8362210B2 (en) | 2010-01-19 | 2013-01-29 | Xencor, Inc. | Antibody variants with enhanced complement activity |
KR20130028055A (en) | 2010-01-28 | 2013-03-18 | 글락소 그룹 리미티드 | Cd127 binding proteins |
EP2534175A2 (en) | 2010-02-09 | 2012-12-19 | Glaxo Group Limited | Treatment of a metabolic disorder |
CA2789629A1 (en) | 2010-02-10 | 2011-08-18 | Immunogen, Inc. | Cd20 antibodies and uses thereof |
WO2011101328A2 (en) | 2010-02-18 | 2011-08-25 | Roche Glycart Ag | Treatment with a humanized igg class anti egfr antibody and an antibody against insulin like growth factor 1 receptor |
JP5981853B2 (en) | 2010-02-18 | 2016-08-31 | ジェネンテック, インコーポレイテッド | Neuregulin antagonists and their use in the treatment of cancer |
EP3696194A1 (en) | 2010-02-23 | 2020-08-19 | F. Hoffmann-La Roche AG | Anti-angiogenesis therapy for the treatment of ovarian cancer |
UA108227C2 (en) | 2010-03-03 | 2015-04-10 | ANTIGENCY PROTEIN | |
KR101899835B1 (en) | 2010-03-24 | 2018-09-19 | 제넨테크, 인크. | Anti-lrp6 antibodies |
EP2374816B1 (en) | 2010-04-07 | 2016-09-28 | Agency For Science, Technology And Research | Binding molecules against Chikungunya virus and uses thereof |
US9441032B2 (en) | 2010-04-07 | 2016-09-13 | Agency For Science, Technology And Research | Binding molecules against Chikungunya virus and uses thereof |
WO2011130332A1 (en) | 2010-04-12 | 2011-10-20 | Academia Sinica | Glycan arrays for high throughput screening of viruses |
JP5947289B2 (en) | 2010-05-10 | 2016-07-06 | アカデミア シニカAcademia Sinica | Determination of the susceptibility of zanamivir phosphonate congeners with anti-influenza activity and influenza virus to oseltamivir |
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 |
UY33421A (en) | 2010-06-03 | 2011-12-30 | Glaxo Wellcome House | HUMANIZED ANTIGEN UNION PROTEINS |
KR20130098165A (en) | 2010-06-03 | 2013-09-04 | 제넨테크, 인크. | Immuno-pet imaging of antibodies and immunoconjugates and uses therefor |
CA2794731C (en) | 2010-06-18 | 2019-03-19 | Genentech, Inc. | Anti-axl antibodies and methods of use |
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 |
SG186983A1 (en) | 2010-07-09 | 2013-02-28 | Genentech Inc | Anti-neuropilin antibodies and methods of use |
EP2409989A1 (en) | 2010-07-19 | 2012-01-25 | International-Drug-Development-Biotech | Method to improve glycosylation profile for antibody |
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 |
EP2596026B1 (en) | 2010-07-23 | 2020-04-08 | Trustees of Boston University | Anti-despr inhibitors as therapeutics for inhibition of pathological angiogenesis and tumor cell invasiveness and for molecular imaging and targeted delivery |
KR20130045914A (en) | 2010-08-03 | 2013-05-06 | 에프. 호프만-라 로슈 아게 | Chronic lymphocytic leukemia (cll) biomarkers |
RU2013106217A (en) | 2010-08-05 | 2014-09-10 | Ф. Хоффманн-Ля Рош Аг | HYBRID PROTEIN FROM ANTIBODIES AGAINST MHC AND ANTIVIRAL CYTOKINE |
RU2584597C2 (en) | 2010-08-13 | 2016-05-20 | Рош Гликарт Аг | Antibodies against a2 tenastin-c and methods for use thereof |
SI2603530T1 (en) | 2010-08-13 | 2018-02-28 | Roche Glycart Ag | Anti-fap antibodies and methods of use |
KR101603001B1 (en) | 2010-08-25 | 2016-03-11 | 에프. 호프만-라 로슈 아게 | Antibodies against il-18r1 and uses thereof |
BR112013004673A8 (en) | 2010-08-31 | 2018-01-02 | Genentech Inc | biomarkers and treatment methods. |
WO2012047968A2 (en) | 2010-10-05 | 2012-04-12 | Genentech, Inc. | Mutant smoothened and methods of using the same |
KR20200059320A (en) | 2010-11-08 | 2020-05-28 | 제넨테크, 인크. | Subcutaneously administered anti-il-6 receptor antibody |
EP2638070B1 (en) | 2010-11-10 | 2016-10-19 | F.Hoffmann-La Roche Ag | Methods and compositions for neural disease immunotherapy |
AU2011333878B2 (en) | 2010-11-23 | 2016-01-14 | Glaxo Group Limited | Antigen binding proteins to oncostatin M (OSM) |
BR112013013003A2 (en) | 2010-11-24 | 2016-08-09 | Glaxo Group Ltd | antigen binding protein and pharmaceutical composition |
NZ706751A (en) | 2010-11-30 | 2016-10-28 | Genentech Inc | Low affinity blood brain barrier receptor antibodies and uses therefor |
AU2011343570B2 (en) | 2010-12-16 | 2016-11-03 | Genentech, Inc. | Diagnosis and treatments relating to TH2 inhibition |
NZ610976A (en) | 2010-12-20 | 2015-07-31 | Genentech Inc | Anti-mesothelin antibodies and immunoconjugates |
JP2014511106A (en) | 2010-12-22 | 2014-05-08 | ジェネンテック, インコーポレイテッド | Anti-PCSK9 antibody and method of use |
WO2012092539A2 (en) | 2010-12-31 | 2012-07-05 | Takeda Pharmaceutical Company Limited | Antibodies to dll4 and uses thereof |
EP2482074A1 (en) * | 2011-01-27 | 2012-08-01 | Medizinische Hochschule Hannover | Methods and means for diagnosing vasculitis |
EP3971206A1 (en) | 2011-02-10 | 2022-03-23 | Roche Glycart AG | Mutant interleukin-2 polypeptides |
BR112013019083A2 (en) | 2011-02-10 | 2017-04-04 | Roche Glycart Ag | combination of (a) an immunoconjugate, pharmaceutical composition, use of (a) an immunoconjugate, method of treating a disease in an individual, method of stimulating cellular function in an individual, and kit for treating a disease. |
EP2681239B8 (en) | 2011-02-28 | 2015-09-09 | F. Hoffmann-La Roche AG | Antigen binding proteins |
BR112013020338A2 (en) | 2011-02-28 | 2016-10-18 | Hoffmann La Roche | monovalent antigen binding protein, pharmaceutical composition, use of monovalent antigen binding protein, method for treating a patient in need of therapy, method for preparing a monovalent antigen binding protein, nucleic acid, vector and cell hostess |
EP2681244B1 (en) | 2011-03-02 | 2017-11-29 | Roche Glycart AG | Cea antibodies |
EP2694551A1 (en) | 2011-04-07 | 2014-02-12 | Genentech, Inc. | Anti-fgfr4 antibodies and methods of use |
EP2702077A2 (en) | 2011-04-27 | 2014-03-05 | AbbVie Inc. | Methods for controlling the galactosylation profile of recombinantly-expressed proteins |
EA201892619A1 (en) | 2011-04-29 | 2019-04-30 | Роше Гликарт Аг | IMMUNOCONJUGATES CONTAINING INTERLEUKIN-2 MUTANT POLYPETIPS |
WO2012146630A1 (en) | 2011-04-29 | 2012-11-01 | F. Hoffmann-La Roche Ag | N-terminal acylated polypeptides, methods for their production and uses thereof |
EP2707723B1 (en) | 2011-05-12 | 2016-02-10 | Genentech, Inc. | Multiple reaction monitoring lc-ms/ms method to detect therapeutic antibodies in animal samples using framework signature pepides |
AU2012255881C1 (en) | 2011-05-16 | 2015-11-26 | Genentech, Inc. | FGFR1 agonists and methods of use |
BR112013028779B8 (en) | 2011-05-27 | 2021-04-20 | Glaxo Group Ltd | antigen-binding protein or immunoconjugate, immunoconjugate, pharmaceutical composition, and, use of a composition |
EP2721067B1 (en) | 2011-06-15 | 2019-07-31 | F.Hoffmann-La Roche Ag | Anti-human epo receptor antibodies and methods of use |
AR086982A1 (en) | 2011-06-22 | 2014-02-05 | Hoffmann La Roche | ELIMINATION OF DIANA CELLS BY SPECIFIC CYTOTOXIC T-CELLS OF VIRUSES USING COMPLEXES THAT INCLUDE MHC CLASS I |
AR086823A1 (en) | 2011-06-30 | 2014-01-22 | Genentech Inc | ANTI-C-MET ANTIBODY FORMULATIONS, METHODS |
US20130022551A1 (en) | 2011-07-22 | 2013-01-24 | Trustees Of Boston University | DEspR ANTAGONISTS AND AGONISTS AS THERAPEUTICS |
BR112014001855A2 (en) | 2011-07-27 | 2017-02-21 | Glaxo Group Ltd | antigen-binding construct and protein, dimer, pharmaceutical composition, polynucleotide sequence, host cell and method for construct production |
CA2842375A1 (en) | 2011-08-17 | 2013-02-21 | Erica Jackson | Neuregulin antibodies and uses thereof |
EP2744825A1 (en) | 2011-08-17 | 2014-06-25 | F.Hoffmann-La Roche Ag | Inhibition of angiogenesis in refractory tumors |
KR20140048292A (en) | 2011-08-23 | 2014-04-23 | 로슈 글리카트 아게 | Anti-mcsp antibodies |
SI2748202T1 (en) | 2011-08-23 | 2018-10-30 | Roche Glycart Ag | Bispecific antigen binding molecules |
BR112014005720A2 (en) | 2011-09-15 | 2017-12-12 | Genentech Inc | method of selecting and / or identifying a usp1 antagonist, uaf1 antagonist and / or an id antagonist that promotes a change in the cellular fate of said method |
RU2014114617A (en) | 2011-09-19 | 2015-10-27 | Дженентек, Инк. | COMBINED TREATMENTS CONTAINING C-MET ANTAGONISTS AND B-RAF ANTAGONISTS |
AU2012319150B2 (en) | 2011-10-05 | 2017-08-17 | Genentech, Inc. | Methods of treating liver conditions using Notch2 antagonists |
RS57645B1 (en) | 2011-10-14 | 2018-11-30 | Hoffmann La Roche | Anti-htra1 antibodies and methods of use |
KR20140077178A (en) | 2011-10-19 | 2014-06-23 | 로슈 글리카트 아게 | Separation method for fucosylated antibodies |
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 |
TW201326193A (en) | 2011-11-21 | 2013-07-01 | Genentech Inc | Purification of anti-c-met antibodies |
EP2788024A1 (en) | 2011-12-06 | 2014-10-15 | F.Hoffmann-La Roche Ag | Antibody formulation |
MX358680B (en) | 2011-12-08 | 2018-08-31 | Biotest Ag | Uses of immunoconjugates targeting cd138. |
KR102229491B1 (en) | 2011-12-22 | 2021-03-18 | 에프. 호프만-라 로슈 아게 | Expression vector element combinations, novel production cell generation methods and their use for the recombinant production of polypeptides |
SG11201403445YA (en) | 2011-12-22 | 2014-07-30 | Hoffmann La Roche | Full length antibody display system for eukaryotic cells and its use |
SG11201403223PA (en) | 2011-12-22 | 2014-07-30 | Hoffmann La Roche | Expression vector organization, novel production cell generation methods and their use for the recombinant production of polypeptides |
WO2013096791A1 (en) | 2011-12-23 | 2013-06-27 | Genentech, Inc. | Process for making high concentration protein formulations |
WO2013101771A2 (en) | 2011-12-30 | 2013-07-04 | Genentech, Inc. | Compositions and method for treating autoimmune diseases |
JP6684490B2 (en) | 2012-01-09 | 2020-04-22 | ザ・スクリップス・リサーチ・インスティテュート | Ultralong complementarity determining regions and uses thereof |
AU2013208007A1 (en) | 2012-01-09 | 2014-07-31 | The Scripps Research Institute | Humanized antibodies with ultralong CDR3 |
WO2013109856A2 (en) | 2012-01-18 | 2013-07-25 | Genentech, Inc. | Methods of using fgf19 modulators |
PE20141561A1 (en) | 2012-01-18 | 2014-11-12 | Genentech Inc | ANTI-LRP5 ANTIBODIES AND METHODS OF USE |
WO2013113641A1 (en) | 2012-01-31 | 2013-08-08 | Roche Glycart Ag | Use of nkp46 as a predictive biomarker for cancer treatment with adcc- enhanced antibodies |
EP2812350B1 (en) | 2012-02-11 | 2019-04-03 | F.Hoffmann-La Roche Ag | R-spondin translocations and methods using the same |
BR112014018005B1 (en) | 2012-02-15 | 2021-06-29 | F. Hoffmann-La Roche Ag | USE OF A NON-COVALENT IMMOBILIZED COMPLEX |
BR112014018374A8 (en) | 2012-03-02 | 2017-07-11 | Roche Glycart Ag | METHOD FOR PREDICTING THE RESPONSE OF A PATIENT WITH CANCER, KIT, ANTIBODY, METHOD FOR THE TREATMENT OF CANCER AND PHARMACEUTICAL COMPOSITION |
US20130259867A1 (en) | 2012-03-27 | 2013-10-03 | Genentech, Inc. | Diagnosis and treatments relating to her3 inhibitors |
AR090549A1 (en) | 2012-03-30 | 2014-11-19 | Genentech Inc | ANTI-LGR5 AND IMMUNOCATE PLAYERS |
US10130714B2 (en) | 2012-04-14 | 2018-11-20 | Academia Sinica | Enhanced anti-influenza agents conjugated with anti-inflammatory activity |
US9067990B2 (en) | 2013-03-14 | 2015-06-30 | Abbvie, Inc. | Protein purification using displacement chromatography |
WO2013158279A1 (en) | 2012-04-20 | 2013-10-24 | Abbvie Inc. | Protein purification methods to reduce acidic species |
US9181572B2 (en) | 2012-04-20 | 2015-11-10 | Abbvie, Inc. | Methods to modulate lysine variant distribution |
TW201402609A (en) | 2012-05-01 | 2014-01-16 | Genentech Inc | Anti-PMEL17 antibodies and immunoconjugates |
WO2013170191A1 (en) | 2012-05-11 | 2013-11-14 | Genentech, Inc. | Methods of using antagonists of nad biosynthesis from nicotinamide |
RU2711089C2 (en) | 2012-05-18 | 2020-01-15 | Дженентек, Инк. | Highly concentrated compositions of monoclonal antibodies |
BR112014028838A2 (en) | 2012-05-21 | 2020-05-12 | Genentech Inc | USES OF AN ANTIBODY, METHOD FOR MAKING AN ANTIBODY AND ANTIBODIES |
DK2852610T3 (en) | 2012-05-23 | 2018-09-03 | Glykos Finland Oy | PRODUCTION OF FUCOSYLED GLYCOPROTEIN |
JP6294311B2 (en) | 2012-05-23 | 2018-03-14 | ジェネンテック, インコーポレイテッド | How to select a treatment |
WO2013176754A1 (en) | 2012-05-24 | 2013-11-28 | Abbvie Inc. | Novel purification of antibodies using hydrophobic interaction chromatography |
CN103463633B (en) * | 2012-06-07 | 2016-03-30 | 复旦大学 | Chimeric hepatitis B virus core antigen therapeutic vaccine of a kind of targeting and uses thereof |
BR112014031310A2 (en) | 2012-06-15 | 2017-07-25 | Genentech Inc | anti-pcsk9 antibodies, formulations, dosage and methods of use |
US20140004121A1 (en) | 2012-06-27 | 2014-01-02 | Amgen Inc. | Anti-mesothelin binding proteins |
CN107082810B (en) | 2012-07-04 | 2020-12-25 | 弗·哈夫曼-拉罗切有限公司 | Anti-theophylline antibodies and methods of use |
WO2014006123A1 (en) | 2012-07-04 | 2014-01-09 | F. Hoffmann-La Roche Ag | Anti-biotin antibodies and methods of use |
EP2869848B1 (en) | 2012-07-04 | 2016-09-21 | F. Hoffmann-La Roche AG | Covalently linked antigen-antibody conjugates |
SI2870247T1 (en) | 2012-07-05 | 2019-10-30 | Hoffmann La Roche | Expression and secretion system |
AU2013288929A1 (en) | 2012-07-09 | 2014-12-04 | Genentech, Inc. | Immunoconjugates comprising anti-CD22 antibodies |
EA201590171A1 (en) | 2012-07-09 | 2015-09-30 | Дженентек, Инк. | IMMUNOCONGATES CONTAINING ANTIBODIES TO CD79b |
SG11201500096YA (en) | 2012-07-09 | 2015-02-27 | Genentech Inc | Immunoconjugates comprising anti - cd79b antibodies |
WO2014011520A1 (en) | 2012-07-09 | 2014-01-16 | Genentech, Inc. | Immunoconjugates comprising anti-cd22 antibodies |
LT2872534T (en) | 2012-07-13 | 2018-10-25 | Roche Glycart Ag | Bispecific anti-vegf/anti-ang-2 antibodies and their use in the treatment of ocular vascular diseases |
NZ630363A (en) | 2012-07-25 | 2018-09-28 | Celldex Therapeutics Inc | Anti-kit antibodies and uses thereof |
CA2872195A1 (en) | 2012-08-07 | 2014-02-13 | Roche Glycart Ag | Composition comprising two antibodies engineered to have reduced and increased effector function |
JP6302909B2 (en) | 2012-08-18 | 2018-03-28 | アカデミア シニカAcademia Sinica | Cell-permeable probes for sialidase identification and imaging |
AU2013305827A1 (en) | 2012-08-21 | 2015-03-05 | Academia Sinica | Benzocyclooctyne compounds and uses thereof |
EP2888279A1 (en) | 2012-08-22 | 2015-07-01 | Glaxo Group Limited | Anti lrp6 antibodies |
SG11201500583PA (en) | 2012-08-29 | 2015-04-29 | Hoffmann La Roche | Blood brain barrier shuttle |
AU2013309506A1 (en) | 2012-09-02 | 2015-03-12 | Abbvie Inc. | Methods to control protein heterogeneity |
US9512214B2 (en) | 2012-09-02 | 2016-12-06 | Abbvie, Inc. | Methods to control protein heterogeneity |
CN111481552A (en) | 2012-09-07 | 2020-08-04 | 吉宁特有限公司 | Combination therapy of type II anti-CD 20 antibodies with selective Bcl-2 inhibitors |
HUE045435T2 (en) | 2012-10-12 | 2019-12-30 | Medimmune Ltd | Pyrrolobenzodiazepines and conjugates thereof |
US10100102B2 (en) | 2012-10-29 | 2018-10-16 | The University Of North Carolina At Chapel Hill | Compositions and methods for inhibiting pathogen infection |
AU2013337277B2 (en) | 2012-11-05 | 2018-03-08 | Foundation Medicine, Inc. | Novel NTRK1 fusion molecules and uses thereof |
WO2014072306A1 (en) | 2012-11-08 | 2014-05-15 | F. Hoffmann-La Roche Ag | Her3 antigen binding proteins binding to the beta-hairpin of her3 |
TWI657095B (en) | 2012-11-13 | 2019-04-21 | 美商建南德克公司 | Anti-hemagglutinin antibodies and methods of use |
WO2014096015A1 (en) | 2012-12-21 | 2014-06-26 | F. Hoffmann-La Roche Ag | Disulfide-linked multivalent mhc class i comprising multi-function proteins |
EP3939614A1 (en) | 2013-01-18 | 2022-01-19 | 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 |
WO2014114595A1 (en) | 2013-01-23 | 2014-07-31 | Roche Glycart Ag | Predictive biomarker for cancer treatment with adcc-enhanced antibodies |
CN105263319A (en) | 2013-02-13 | 2016-01-20 | 法国化学与生物科技实验室 | Proteins with modified glycosylation and methods of production thereof |
TW201506041A (en) | 2013-02-13 | 2015-02-16 | Lab Francais Du Fractionnement | highly galactosylated anti-TNF-alpha antibodies and uses thereof |
KR20150118159A (en) | 2013-02-22 | 2015-10-21 | 에프. 호프만-라 로슈 아게 | Methods of treating cancer and preventing drug resistance |
CN104936987A (en) | 2013-02-26 | 2015-09-23 | 罗切格利卡特公司 | Anti-MCSP antibodies |
US9925240B2 (en) | 2013-03-06 | 2018-03-27 | Genentech, Inc. | Methods of treating and preventing cancer drug resistance |
CA2905010A1 (en) | 2013-03-12 | 2014-09-18 | Abbvie Inc. | Human antibodies that bind human tnf-alpha and methods of preparing the same |
WO2014140174A1 (en) | 2013-03-13 | 2014-09-18 | Spirogen Sàrl | Pyrrolobenzodiazepines and conjugates thereof |
US9017687B1 (en) | 2013-10-18 | 2015-04-28 | Abbvie, Inc. | Low acidic species compositions and methods for producing and using the same using displacement chromatography |
JP2016515132A (en) | 2013-03-14 | 2016-05-26 | ジェネンテック, インコーポレイテッド | Combination and use of MEK inhibitor compounds with HER3 / EGFR inhibitor compounds |
US8921526B2 (en) | 2013-03-14 | 2014-12-30 | Abbvie, Inc. | Mutated anti-TNFα antibodies and methods of their use |
US9499614B2 (en) | 2013-03-14 | 2016-11-22 | Abbvie Inc. | Methods for modulating protein glycosylation profiles of recombinant protein therapeutics using monosaccharides and oligosaccharides |
US9562099B2 (en) | 2013-03-14 | 2017-02-07 | Genentech, Inc. | Anti-B7-H4 antibodies and immunoconjugates |
EP3299391B1 (en) | 2013-03-14 | 2019-12-04 | Genentech, Inc. | Anti-b7-h4 antibodies and immunoconjugates |
MX2015011606A (en) | 2013-03-14 | 2016-05-17 | Genentech Inc | Methods of treating cancer and preventing cancer drug resistance. |
MX368005B (en) | 2013-03-15 | 2019-09-13 | Genentech Inc | Il-22 polypeptides and il-22 fc fusion proteins and methods of use. |
MX2015011899A (en) | 2013-03-15 | 2016-05-05 | Genentech Inc | Methods of treating cancer and preventing cancer drug resistance. |
US10344088B2 (en) | 2013-03-15 | 2019-07-09 | Glaxosmithkline Intellectual Property Development Limited | Antigen binding proteins |
AU2014235453A1 (en) | 2013-03-15 | 2015-10-08 | Genentech, Inc. | Biomarkers and methods of treating PD-1 and PD-L1 related conditions |
JP6527132B2 (en) | 2013-03-15 | 2019-06-05 | ジェネンテック, インコーポレイテッド | Compositions and methods for diagnosis and treatment of liver cancer |
WO2014150877A2 (en) | 2013-03-15 | 2014-09-25 | Ac Immune S.A. | Anti-tau antibodies and methods of use |
JP2016517441A (en) | 2013-03-15 | 2016-06-16 | ジェネンテック, インコーポレイテッド | Anti-CRTh2 antibody and method of use |
UA118028C2 (en) | 2013-04-03 | 2018-11-12 | Рош Глікарт Аг | Bispecific antibodies specific for fap and dr5, antibodies specific for dr5 and methods of use |
EP2992010B1 (en) | 2013-04-29 | 2021-03-24 | F.Hoffmann-La Roche Ag | Fc-receptor binding modified asymmetric antibodies and methods of use |
AU2014261630B2 (en) | 2013-04-29 | 2019-05-09 | F. Hoffmann-La Roche Ag | Human FcRn-binding modified antibodies and methods of use |
AU2014261631B2 (en) | 2013-04-29 | 2019-02-14 | F. Hoffmann-La Roche Ag | FcRn-binding abolished anti-IGF-1R antibodies and their use in the treatment of vascular eye diseases |
KR102293064B1 (en) | 2013-05-20 | 2021-08-23 | 제넨테크, 인크. | Anti-transferrin receptor antibodies and methods of use |
WO2014210397A1 (en) | 2013-06-26 | 2014-12-31 | Academia Sinica | Rm2 antigens and use thereof |
US9981030B2 (en) | 2013-06-27 | 2018-05-29 | Academia Sinica | Glycan conjugates and use thereof |
WO2015017146A2 (en) | 2013-07-18 | 2015-02-05 | Fabrus, Inc. | Antibodies with ultralong complementarity determining regions |
WO2015010100A2 (en) | 2013-07-18 | 2015-01-22 | Fabrus, Inc. | Humanized antibodies with ultralong complementarity determining regions |
PT3708583T (en) | 2013-08-01 | 2022-05-13 | Five Prime Therapeutics Inc | Afucosylated anti-fgfr2iiib antibodies |
US10093978B2 (en) | 2013-08-12 | 2018-10-09 | Genentech, Inc. | Compositions for detecting complement factor H (CFH) and complement factor I (CFI) polymorphisms |
CN105682666B (en) | 2013-09-06 | 2021-06-01 | 中央研究院 | Activation of human iNKT cells using glycolipids |
CN105518027A (en) | 2013-09-17 | 2016-04-20 | 豪夫迈·罗氏有限公司 | Methods of using anti-LGR5 antibodies |
EP3052640A2 (en) | 2013-10-04 | 2016-08-10 | AbbVie Inc. | Use of metal ions for modulation of protein glycosylation profiles of recombinant proteins |
EP3055328A1 (en) | 2013-10-11 | 2016-08-17 | F. Hoffmann-La Roche AG | Nsp4 inhibitors and methods of use |
JP6502931B2 (en) | 2013-10-11 | 2019-04-17 | アメリカ合衆国 | TEM 8 antibody and use thereof |
CA2922912A1 (en) | 2013-10-11 | 2015-04-16 | F. Hoffmann-La Roche Ag | Multispecific domain exchanged common variable light chain antibodies |
CA2925598A1 (en) | 2013-10-18 | 2015-04-23 | Genentech, Inc. | Anti-rspo antibodies and methods of use |
US8946395B1 (en) | 2013-10-18 | 2015-02-03 | Abbvie Inc. | Purification of proteins using hydrophobic interaction chromatography |
US9181337B2 (en) | 2013-10-18 | 2015-11-10 | Abbvie, Inc. | Modulated lysine variant species compositions and methods for producing and using the same |
US9085618B2 (en) | 2013-10-18 | 2015-07-21 | Abbvie, Inc. | Low acidic species compositions and methods for producing and using the same |
SG11201603127WA (en) | 2013-10-23 | 2016-05-30 | Genentech Inc | Methods of diagnosing and treating eosinophilic disorders |
WO2015073884A2 (en) | 2013-11-15 | 2015-05-21 | Abbvie, Inc. | Glycoengineered binding protein compositions |
MX2016005631A (en) | 2013-11-21 | 2016-07-14 | Hoffmann La Roche | ANTI-alpha-SYNUCLEIN ANTIBODIES AND METHODS OF USE. |
AU2014363944B2 (en) | 2013-12-09 | 2020-03-26 | Allakos Inc. | Anti-Siglec-8 antibodies and methods of use thereof |
EP3080611B1 (en) | 2013-12-13 | 2018-11-14 | The General Hospital Corporation | Soluble high molecular weight (hmw) tau species and applications thereof |
EP3080164B1 (en) | 2013-12-13 | 2019-01-16 | Genentech, Inc. | Anti-cd33 antibodies and immunoconjugates |
HUE050156T2 (en) | 2013-12-17 | 2020-11-30 | Genentech Inc | Anti-cd3 antibodies and methods of use |
MX2016007972A (en) | 2013-12-17 | 2016-10-28 | Genentech Inc | Methods of treating cancers using pd-1 axis binding antagonists and taxanes. |
RU2016128726A (en) | 2013-12-17 | 2018-01-23 | Дженентек, Инк. | METHODS FOR TREATING MALIGNANT TUMORS USING PD-1 BINDING ANTAGONISTS AND ANTIBODIES AGAINST CD20 |
US20150190506A1 (en) | 2013-12-17 | 2015-07-09 | Genentech, Inc. | Combination therapy comprising ox40 binding agonists and pd-1 axis binding antagonists |
TWI728373B (en) | 2013-12-23 | 2021-05-21 | 美商建南德克公司 | Antibodies and methods of use |
ES2742682T3 (en) | 2013-12-24 | 2020-02-17 | Argenx Bvba | FCRN antagonists and methods of use |
JP6476194B2 (en) | 2014-01-03 | 2019-02-27 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Bispecific anti-hapten / anti-blood brain barrier receptor antibodies, complexes thereof, and their use as blood brain barrier shuttles |
CN105873616B (en) | 2014-01-03 | 2020-06-05 | 豪夫迈·罗氏有限公司 | Covalently linked polypeptide toxin-antibody conjugates |
RU2694981C2 (en) | 2014-01-03 | 2019-07-18 | Ф. Хоффманн-Ля Рош Аг | Covalently linked conjugates chelicar-antibody against chelicar and use thereof |
WO2015103549A1 (en) | 2014-01-03 | 2015-07-09 | The United States Of America, As Represented By The Secretary Department Of Health And Human Services | Neutralizing antibodies to hiv-1 env and their use |
RU2694659C2 (en) | 2014-01-06 | 2019-07-16 | Ф. Хоффманн-Ля Рош Аг | Monovalent carrier modules across blood-brain barrier |
RU2727639C2 (en) | 2014-01-15 | 2020-07-22 | Ф.Хоффманн-Ля Рош Аг | Variants of fc-region with modified ability to bind to fcrn and with preserved ability to bind with protein a |
US10150818B2 (en) | 2014-01-16 | 2018-12-11 | Academia Sinica | Compositions and methods for treatment and detection of cancers |
EP3094352B1 (en) | 2014-01-16 | 2020-09-23 | Academia Sinica | Compositions and methods for treatment and detection of cancers |
EP3096797A1 (en) | 2014-01-24 | 2016-11-30 | F. Hoffmann-La Roche AG | Methods of using anti-steap1 antibodies and immunoconjugates |
JP6736467B2 (en) | 2014-02-04 | 2020-08-05 | ジェネンテック, インコーポレイテッド | Smoothing mutant and method of using the same |
WO2015120280A1 (en) | 2014-02-08 | 2015-08-13 | Genentech, Inc. | Methods of treating alzheimer's disease |
SG11201606316XA (en) | 2014-02-08 | 2016-08-30 | Genentech Inc | Methods of treating alzheimer's disease |
TW201902515A (en) | 2014-02-12 | 2019-01-16 | 美商建南德克公司 | 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 |
US10183996B2 (en) | 2014-02-28 | 2019-01-22 | Allakos Inc. | Methods and compositions for treating Siglec-8 associated diseases |
WO2015139046A1 (en) | 2014-03-14 | 2015-09-17 | Genentech, Inc. | Methods and compositions for secretion of heterologous polypeptides |
US20170107294A1 (en) | 2014-03-21 | 2017-04-20 | Nordlandssykehuset Hf | Anti-cd14 antibodies and uses thereof |
MA39776A (en) | 2014-03-24 | 2017-02-01 | Hoffmann La Roche | Cancer treatment with c-met antagonists and correlation of the latter with hgf expression |
EP3129767B1 (en) | 2014-03-27 | 2021-09-01 | Academia Sinica | Reactive labelling compounds and uses thereof |
MA39817A (en) | 2014-03-31 | 2017-02-08 | Hoffmann La Roche | Combination therapy comprising anti-angiogenesis agents and ox40 binding agonists |
EP3632934A1 (en) | 2014-03-31 | 2020-04-08 | F. Hoffmann-La Roche AG | Anti-ox40 antibodies and methods of use |
WO2015164615A1 (en) | 2014-04-24 | 2015-10-29 | University Of Oslo | Anti-gluten antibodies and uses thereof |
CN106414499A (en) | 2014-05-22 | 2017-02-15 | 基因泰克公司 | Anti-GPC3 antibodies and immunoconjugates |
RU2016144405A (en) | 2014-05-23 | 2018-06-26 | Дженентек, Инк. | MiT BIOMARKERS AND WAYS OF THEIR APPLICATION |
TWI717319B (en) | 2014-05-27 | 2021-02-01 | 中央研究院 | Fucosidase from bacteroides and methods using the same |
US10118969B2 (en) | 2014-05-27 | 2018-11-06 | Academia Sinica | Compositions and methods relating to universal glycoforms for enhanced antibody efficacy |
TWI679020B (en) | 2014-05-27 | 2019-12-11 | 中央研究院 | Anti-her2 glycoantibodies and uses thereof |
KR20170003720A (en) | 2014-05-27 | 2017-01-09 | 아카데미아 시니카 | Anti-cd20 glycoantibodies and uses thereof |
KR102494193B1 (en) | 2014-05-28 | 2023-01-31 | 아카데미아 시니카 | Anti-tnf-alpha glycoantibodies and uses thereof |
EP3155015A1 (en) | 2014-06-11 | 2017-04-19 | F. Hoffmann-La Roche AG | Anti-lgr5 antibodies and uses thereof |
CN107073121A (en) | 2014-06-13 | 2017-08-18 | 基因泰克公司 | Treatment and the method for prevention cancer drug resistance |
WO2015197736A1 (en) | 2014-06-26 | 2015-12-30 | F. Hoffmann-La Roche Ag | Anti-brdu antibodies and methods of use |
BR112017000130A2 (en) | 2014-07-11 | 2018-01-09 | Genentech Inc | method for mitigating toxicity associated with notch pathway inhibition and cancer treatment method |
EP3309174B1 (en) | 2014-07-11 | 2022-05-11 | Ventana Medical Systems, Inc. | Anti-pd-l1 antibodies and diagnostic uses thereof |
CN108064266A (en) | 2014-07-21 | 2018-05-22 | 格利科斯芬兰公司 | The preparation of the glycoprotein with mammal sample N- glycan in filamentous fungi |
TWI790593B (en) | 2014-08-19 | 2023-01-21 | 美商默沙東有限責任公司 | Anti-tigit antibodies |
AU2015308818B2 (en) | 2014-08-28 | 2021-02-25 | Bioatla Llc | Conditionally active chimeric antigen receptors for modified T-cells |
TWI805109B (en) | 2014-08-28 | 2023-06-11 | 美商奇諾治療有限公司 | Antibodies and chimeric antigen receptors specific for cd19 |
CN107001404B (en) | 2014-09-08 | 2021-06-29 | 中央研究院 | Activation of human iNKT cells using glycolipids |
JP6531166B2 (en) | 2014-09-10 | 2019-06-12 | メドイミューン・リミテッドMedImmune Limited | Pyrrolobenzodiazepine and its conjugate |
JP6886398B2 (en) | 2014-09-12 | 2021-06-16 | ジェネンテック, インコーポレイテッド | ANTI-CLL-1 antibody and immune complex |
SG11201701623UA (en) | 2014-09-12 | 2017-03-30 | Genentech Inc | Anti-her2 antibodies and immunoconjugates |
CN113698485A (en) | 2014-09-12 | 2021-11-26 | 基因泰克公司 | anti-B7-H4 antibodies and immunoconjugates |
RU2727663C2 (en) | 2014-09-17 | 2020-07-22 | Дженентек, Инк. | Immunoconjugates, containing antibodies against her2 and pyrrolbenzodiazepines |
EP3689910A3 (en) | 2014-09-23 | 2020-12-02 | F. Hoffmann-La Roche AG | Method of using anti-cd79b immunoconjugates |
MA40764A (en) | 2014-09-26 | 2017-08-01 | Chugai Pharmaceutical Co Ltd | THERAPEUTIC AGENT INDUCING CYTOTOXICITY |
EP3207057A2 (en) | 2014-10-16 | 2017-08-23 | F. Hoffmann-La Roche AG | Anti-alpha-synuclein antibodies and methods of use |
WO2016059602A2 (en) | 2014-10-16 | 2016-04-21 | Glaxo Group Limited | Methods of treating cancer and related compositions |
EP3223865A4 (en) | 2014-10-31 | 2018-10-03 | Jounce Therapeutics, Inc. | Methods of treating conditions with antibodies that bind b7-h4 |
AU2015343337A1 (en) | 2014-11-03 | 2017-06-15 | Genentech, Inc. | Assays for detecting T cell immune subsets and methods of use thereof |
EP3215637B1 (en) | 2014-11-03 | 2019-07-03 | F. Hoffmann-La Roche AG | Methods and biomarkers for predicting efficacy and valuation of an ox40 agonist treatment |
US10208120B2 (en) | 2014-11-05 | 2019-02-19 | Genentech, Inc. | Anti-FGFR2/3 antibodies and methods using same |
JP6576456B2 (en) | 2014-11-06 | 2019-09-18 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Fc region variants with modified FcRn binding properties and protein A binding properties |
EP3215536A1 (en) | 2014-11-06 | 2017-09-13 | F. Hoffmann-La Roche AG | Combination therapy comprising ox40 binding agonists and tigit inhibitors |
RU2714116C2 (en) | 2014-11-06 | 2020-02-11 | Ф. Хоффманн-Ля Рош Аг | VARIANTS OF Fc-DOMAIN WITH MODIFIED FcRn BINDING AND METHODS OF APPLICATION THEREOF |
JP6929771B2 (en) | 2014-11-10 | 2021-09-01 | ジェネンテック, インコーポレイテッド | Anti-interleukin-33 antibody and its use |
CN107105632A (en) | 2014-11-10 | 2017-08-29 | 豪夫迈·罗氏有限公司 | Nephrosis animal model and its therapeutic agent |
US10160795B2 (en) | 2014-11-14 | 2018-12-25 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Neutralizing antibodies to Ebola virus glycoprotein and their use |
BR112017009006A2 (en) | 2014-11-14 | 2018-04-10 | F. Hoffmann-La Roche Ag | binding molecule, isolated polynucleotide, vector, host cell, binding molecule production method, pharmaceutical composition, use of the binding molecule, and method of treating a disease in an individual |
SG10201807625PA (en) | 2014-11-17 | 2018-10-30 | Genentech Inc | Combination therapy comprising ox40 binding agonists and pd-1 axis binding antagonists |
LT3221349T (en) | 2014-11-19 | 2021-02-10 | Axon Neuroscience Se | Humanized tau antibodies in alzheimer's disease |
US10508151B2 (en) | 2014-11-19 | 2019-12-17 | Genentech, Inc. | Anti-transferrin receptor antibodies and methods of use |
JP6993228B2 (en) | 2014-11-19 | 2022-03-03 | ジェネンテック, インコーポレイテッド | Anti-transferrin receptor / anti-BACE1 multispecific antibody and usage |
US10882920B2 (en) | 2014-11-19 | 2021-01-05 | Genentech, Inc. | Antibodies against BACE1 and use thereof for neural disease immunotherapy |
MY192999A (en) | 2014-11-20 | 2022-09-20 | Hoffmann La Roche | Combination therapy of t cell activating bispecific antigen binding molecules and pd-1 axis binding antagonists |
WO2016087416A1 (en) | 2014-12-03 | 2016-06-09 | F. Hoffmann-La Roche Ag | Multispecific antibodies |
DK3227336T3 (en) | 2014-12-05 | 2019-09-16 | Hoffmann La Roche | Anti-CD79b antibodies and methods for use |
EP3230317A2 (en) | 2014-12-10 | 2017-10-18 | F. Hoffmann-La Roche AG | Blood brain barrier receptor antibodies and methods of use |
EP3233921B1 (en) | 2014-12-19 | 2021-09-29 | Chugai Seiyaku Kabushiki Kaisha | Anti-c5 antibodies and methods of use |
US20160200815A1 (en) | 2015-01-05 | 2016-07-14 | Jounce Therapeutics, Inc. | Antibodies that inhibit tim-3:lilrb2 interactions and uses thereof |
CA2973964A1 (en) | 2015-01-16 | 2016-07-21 | Juno Therapeutics, Inc. | Antibodies and chimeric antigen receptors specific for ror1 |
US9975965B2 (en) | 2015-01-16 | 2018-05-22 | Academia Sinica | Compositions and methods for treatment and detection of cancers |
US10495645B2 (en) | 2015-01-16 | 2019-12-03 | Academia Sinica | Cancer markers and methods of use thereof |
EP3247723A1 (en) | 2015-01-22 | 2017-11-29 | Chugai Seiyaku Kabushiki Kaisha | A combination of two or more anti-c5 antibodies and methods of use |
JP6779887B2 (en) | 2015-01-24 | 2020-11-04 | アカデミア シニカAcademia Sinica | New glycan conjugate and how to use it |
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 |
CA2975875A1 (en) | 2015-02-04 | 2016-08-11 | Genentech, Inc. | Mutant smoothened and methods of using the same |
SG10201907215QA (en) | 2015-02-05 | 2019-09-27 | Chugai Pharmaceutical Co Ltd | Antibodies Comprising An Ion Concentration Dependent Antigen-Binding Domain, Fc Region Variants, Il-8-Binding Antibodies, And Uses Therof |
KR20170140180A (en) | 2015-02-24 | 2017-12-20 | 더 유나이티드 스테이츠 오브 어메리카, 애즈 리프리젠티드 바이 더 세크러테리, 디파트먼트 오브 헬쓰 앤드 휴먼 서비씨즈 | Middle east respiratory syndrome coronavirus immunogens, antibodies, and their use |
KR20170131463A (en) | 2015-03-09 | 2017-11-29 | 아르제넥스 비브이비에이 | Methods of Reducing Serum Levels of Fc-Containing Agents Using FcRn Antagonists |
CA2977285A1 (en) | 2015-03-16 | 2016-09-22 | 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 |
AU2016235541B2 (en) | 2015-03-20 | 2021-04-01 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Neutralizing antibodies to gp120 and their use |
CN107743495B (en) | 2015-03-23 | 2021-05-14 | 拜耳制药股份公司 | anti-CEACAM 6 antibodies and uses thereof |
WO2016154177A2 (en) | 2015-03-23 | 2016-09-29 | Jounce Therapeutics, Inc. | Antibodies to icos |
SG11201707490SA (en) | 2015-04-03 | 2017-10-30 | Eureka Therapeutics Inc | Constructs targeting afp peptide/mhc complexes and uses thereof |
ES2881694T3 (en) | 2015-04-24 | 2021-11-30 | Hoffmann La Roche | Procedures for Identifying Bacteria Comprising Binding 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 |
AU2016256911B2 (en) | 2015-05-07 | 2022-03-31 | Agenus Inc. | Anti-OX40 antibodies and methods of use thereof |
JP6963508B2 (en) | 2015-05-11 | 2021-11-10 | ジェネンテック, インコーポレイテッド | Compositions and Methods for Treating Lupus Nephritis |
CA2983282A1 (en) | 2015-05-12 | 2016-11-17 | Genentech, Inc. | Therapeutic and diagnostic methods for cancer |
WO2016196381A1 (en) | 2015-05-29 | 2016-12-08 | Genentech, Inc. | Pd-l1 promoter methylation in cancer |
JP2018520658A (en) | 2015-05-29 | 2018-08-02 | ジェネンテック, インコーポレイテッド | Humanized anti-Ebola virus glycoprotein antibodies and uses thereof |
CA2984003A1 (en) | 2015-05-29 | 2016-12-08 | Genentech, Inc. | Therapeutic and diagnostic methods for cancer |
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 |
WO2016196975A1 (en) | 2015-06-03 | 2016-12-08 | The United States Of America, As Represented By The Secretary Department Of Health & Human Services | Neutralizing antibodies to hiv-1 env and their use |
JP6360265B2 (en) | 2015-06-04 | 2018-07-18 | オスペダーレ・サン・ラッファエーレ・エッセエッレエッレ | IGFBP3 / TMEM219 axis and diabetes inhibitor |
US10682391B2 (en) | 2015-06-04 | 2020-06-16 | Ospedale San Raffaele Srl | Inhibitors of IGFBP3 binding to TMEM219 for treatment of intestinal diseases |
CA2986942A1 (en) | 2015-06-05 | 2016-12-08 | Genentech, Inc. | Anti-tau antibodies and methods of use |
KR20180011839A (en) | 2015-06-08 | 2018-02-02 | 제넨테크, 인크. | Treatment of Cancer Using Anti-OX40 Antibody |
US20170000885A1 (en) | 2015-06-08 | 2017-01-05 | Genentech, Inc. | Methods of treating cancer using anti-ox40 antibodies and pd-1 axis binding antagonists |
CN108064246A (en) | 2015-06-15 | 2018-05-22 | 基因泰克公司 | Antibody and immune conjugate |
HRP20231134T1 (en) | 2015-06-16 | 2024-01-05 | F. Hoffmann - La Roche Ag | Humanized and affinity matured antibodies to fcrh5 and methods of use |
JP2018526972A (en) | 2015-06-16 | 2018-09-20 | ジェネンテック, インコーポレイテッド | Anti-CD3 antibody and method 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 |
AU2016280159A1 (en) | 2015-06-17 | 2017-12-07 | Genentech, Inc. | Anti-HER2 antibodies and methods of use |
AU2016280070B2 (en) | 2015-06-17 | 2022-09-15 | Genentech, Inc. | Methods of treating locally advanced or metastatic breast cancers using PD-1 axis binding antagonists and taxanes |
AU2016278239B9 (en) | 2015-06-17 | 2022-08-11 | Allakos Inc. | Methods and compositions for treating fibrotic diseases |
HRP20220304T1 (en) | 2015-06-24 | 2022-05-13 | F. Hoffmann - La Roche Ag | Anti-transferrin receptor antibodies with tailored affinity |
BR112017027736A2 (en) | 2015-06-29 | 2018-10-09 | Genentech Inc | anti-cd20 type ii antibody for use in organ transplantation |
WO2017001350A1 (en) | 2015-06-29 | 2017-01-05 | Ventana Medical Systems, Inc. | Materials and methods for performing histochemical assays for human pro-epiregulin and amphiregulin |
RU2611685C2 (en) * | 2015-07-20 | 2017-02-28 | Илья Владимирович Духовлинов | Humanized monoclonal antibody specific to syndecan-1 |
AU2016304764C1 (en) | 2015-08-07 | 2023-06-01 | Imaginab, Inc. | Antigen binding constructs to target molecules |
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 |
EP3341415B1 (en) | 2015-08-28 | 2021-03-24 | H. Hoffnabb-La Roche Ag | Anti-hypusine antibodies and uses thereof |
RU2760582C2 (en) | 2015-09-02 | 2021-11-29 | Иммутеп С.А.С. | Anti-lag-3 antibodies |
CA2999369C (en) | 2015-09-22 | 2023-11-07 | Spring Bioscience Corporation | Anti-ox40 antibodies and diagnostic uses thereof |
PE20181363A1 (en) | 2015-09-23 | 2018-08-27 | Genentech Inc | OPTIMIZED VARIANTS OF ANTI-VEGF ANTIBODIES |
US11142565B2 (en) | 2015-09-24 | 2021-10-12 | Abvitro Llc | Broadly neutralizing anti-HIV-1 antibodies that bind to an N-glycan epitope on the envelope |
EP3356415A1 (en) | 2015-09-29 | 2018-08-08 | Amgen Inc. | Asgr inhibitors |
AR106188A1 (en) | 2015-10-01 | 2017-12-20 | Hoffmann La Roche | ANTI-CD19 HUMANIZED HUMAN ANTIBODIES AND METHODS OF USE |
EP3150636A1 (en) | 2015-10-02 | 2017-04-05 | F. Hoffmann-La Roche AG | Tetravalent multispecific antibodies |
LT3356411T (en) | 2015-10-02 | 2021-09-10 | F. Hoffmann-La Roche Ag | Bispecific antibodies specific for pd1 and tim3 |
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 |
EP3356406A1 (en) | 2015-10-02 | 2018-08-08 | H. Hoffnabb-La Roche Ag | Bispecific anti-human cd20/human transferrin receptor antibodies and methods of use |
MA43345A (en) | 2015-10-02 | 2018-08-08 | Hoffmann La Roche | PYRROLOBENZODIAZEPINE ANTIBODY-DRUG CONJUGATES AND METHODS OF USE |
RU2761115C1 (en) | 2015-10-02 | 2021-12-06 | Ф. Хоффманн-Ля Рош Аг | Bispecific antibodies specific relatively to costimulatory tnf-receptor |
EP3359572A2 (en) | 2015-10-06 | 2018-08-15 | H. Hoffnabb-La Roche Ag | Method for treating multiple sclerosis |
US10392441B2 (en) | 2015-10-07 | 2019-08-27 | United States Of America, As Represented By The Secretary, Department Of Health And Human Services | IL-7R-alpha specific antibodies for treating acute lymphoblastic leukemia |
MY193013A (en) | 2015-10-07 | 2022-09-22 | Hoffmann La Roche | Bispecific antibodies with tetravalency for a costimulatory tnf receptor |
KR20180066236A (en) | 2015-10-22 | 2018-06-18 | 조운스 테라퓨틱스, 인크. | Gene traits for measuring ICOS expression |
US10604577B2 (en) | 2015-10-22 | 2020-03-31 | Allakos Inc. | Methods and compositions for treating systemic mastocytosis |
JO3555B1 (en) | 2015-10-29 | 2020-07-05 | Merck Sharp & Dohme | Antibody neutralizing human respiratory syncytial virus |
EP3184547A1 (en) | 2015-10-29 | 2017-06-28 | F. Hoffmann-La Roche AG | Anti-tpbg antibodies and methods of use |
IL295756A (en) | 2015-10-29 | 2022-10-01 | Hoffmann La Roche | Anti-variant fc-region antibodies and methods of use |
TW201730211A (en) | 2015-10-30 | 2017-09-01 | 建南德克公司 | Anti-Factor D antibodies and conjugates |
PE20181009A1 (en) | 2015-10-30 | 2018-06-26 | Genentech Inc | ANTI-HtrA1 ANTIBODIES AND METHODS OF USE OF THEM |
EP4011911A1 (en) | 2015-11-03 | 2022-06-15 | The United States of America as represented by The Secretary Department of Health and Human Services | Neutralizing antibodies to hiv-1 gp41 and their use |
CN108602884A (en) | 2015-11-08 | 2018-09-28 | 豪夫迈·罗氏有限公司 | The method for screening multi-specificity antibody |
TWI791422B (en) | 2015-11-23 | 2023-02-11 | 美商戊瑞治療有限公司 | Fgfr2 inhibitors alone or in combination with immune stimulating agents in cancer treatment |
JP7325186B2 (en) | 2015-12-09 | 2023-08-14 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Type II anti-CD20 antibody for reducing the formation of anti-drug antibodies |
TWI597292B (en) | 2015-12-18 | 2017-09-01 | 中外製藥股份有限公司 | Anti-c5 antibodies and methods of use |
WO2017117311A1 (en) | 2015-12-30 | 2017-07-06 | Genentech, Inc. | Formulations with reduced degradation of polysorbate |
PL3400246T3 (en) | 2016-01-08 | 2021-03-08 | F. Hoffmann-La Roche Ag | Methods of treating cea-positive cancers using pd-1 axis binding antagonists and anti-cea/anti-cd3 bispecific antibodies |
CA3011739A1 (en) | 2016-01-20 | 2017-07-27 | Genentech, Inc. | High dose treatments for alzheimer's disease |
AU2017212484C1 (en) | 2016-01-27 | 2020-11-05 | Medimmune, Llc | Methods for preparing antibodies with a defined glycosylation pattern |
WO2017136558A1 (en) | 2016-02-04 | 2017-08-10 | Curis, Inc. | Mutant smoothened and methods of using the same |
WO2017145166A1 (en) | 2016-02-25 | 2017-08-31 | B. G. Negev Technologies And Applications Ltd., At Ben-Gurion University | Composition and method for treating amyotrophic lateral sclerosis |
CN114395624A (en) | 2016-02-29 | 2022-04-26 | 基因泰克公司 | Methods for treatment and diagnosis of cancer |
CA3016170A1 (en) | 2016-03-08 | 2017-09-14 | Academia Sinica | Methods for modular synthesis of n-glycans and arrays thereof |
MX2018010546A (en) | 2016-03-15 | 2019-02-20 | Chugai Pharmaceutical Co Ltd | Methods of treating cancers using pd-1 axis binding antagonists and anti-gpc3 antibodies. |
JP6943872B2 (en) | 2016-03-25 | 2021-10-06 | ジェネンテック, インコーポレイテッド | Multiple whole antibody and antibody complex drug quantification assay |
WO2017172771A2 (en) | 2016-03-29 | 2017-10-05 | Janssen Biotech, Inc. | Method of treating psoriasis with increased interval dosing of anti-il12/23 antibody |
BR112018070534A2 (en) | 2016-04-05 | 2019-02-12 | Glaxosmithkline Intellectual Property Development Limited | tgfbeta inhibition in immunotherapy |
EP3443004A1 (en) | 2016-04-14 | 2019-02-20 | H. Hoffnabb-La Roche Ag | Anti-rspo3 antibodies and methods of use |
CN109154613A (en) | 2016-04-15 | 2019-01-04 | 豪夫迈·罗氏有限公司 | For monitoring and the method for the treatment of cancer |
EP3443120A2 (en) | 2016-04-15 | 2019-02-20 | H. Hoffnabb-La Roche Ag | Methods for monitoring and treating cancer |
SG11201808994YA (en) | 2016-04-15 | 2018-11-29 | Bioatla Llc | Anti-axl antibodies, antibody fragments and their immunoconjugates and uses thereof |
WO2017192589A1 (en) | 2016-05-02 | 2017-11-09 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Neutralizing antibodies to influenza ha and their use and identification |
UA123323C2 (en) | 2016-05-02 | 2021-03-17 | Ф. Хоффманн-Ля Рош Аг | The contorsbody - a single chain target binder |
EP3455252B1 (en) | 2016-05-11 | 2022-02-23 | F. Hoffmann-La Roche AG | Modified anti-tenascin antibodies and methods of use |
JP7285076B2 (en) | 2016-05-11 | 2023-06-01 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Antigen-binding molecule comprising a TNF family ligand trimer and a tenascin-binding portion |
EP3243836A1 (en) | 2016-05-11 | 2017-11-15 | F. Hoffmann-La Roche AG | C-terminally fused tnf family ligand trimer-containing antigen binding molecules |
IL308504A (en) | 2016-05-13 | 2024-01-01 | Bioatla Llc | Anti-ror2 antibodies, antibody fragments, their immunoconjugates and uses thereof |
EP3243832A1 (en) | 2016-05-13 | 2017-11-15 | F. Hoffmann-La Roche AG | Antigen binding molecules comprising a tnf family ligand trimer and pd1 binding moiety |
PL3458101T3 (en) | 2016-05-20 | 2021-05-31 | F. Hoffmann-La Roche Ag | Protac antibody conjugates and methods of use |
US20170370906A1 (en) | 2016-05-27 | 2017-12-28 | Genentech, Inc. | Bioanalytical analysis of site-specific antibody drug conjugates |
EP3252078A1 (en) | 2016-06-02 | 2017-12-06 | F. Hoffmann-La Roche AG | Type ii anti-cd20 antibody and anti-cd20/cd3 bispecific antibody for treatment of cancer |
CN109563160B (en) | 2016-06-24 | 2023-02-28 | 豪夫迈·罗氏有限公司 | Anti-polyubiquitin multispecific antibodies |
WO2018007314A1 (en) | 2016-07-04 | 2018-01-11 | F. Hoffmann-La Roche Ag | Novel antibody format |
WO2018014260A1 (en) | 2016-07-20 | 2018-01-25 | Nanjing Legend Biotech Co., Ltd. | Multispecific antigen binding proteins and methods of use thereof |
AU2017303205A1 (en) | 2016-07-29 | 2019-01-17 | Chugai Seiyaku Kabushiki Kaisha | Bispecific antibody exhibiting increased alternative FVIII-cofactor-function activity |
MX2019001184A (en) | 2016-07-29 | 2019-09-26 | Juno Therapeutics Inc | Anti-idiotypic antibodies against anti-cd19 antibodies. |
NL2017270B1 (en) | 2016-08-02 | 2018-02-09 | Aduro Biotech Holdings Europe B V | New anti-hCTLA-4 antibodies |
CN116271014A (en) | 2016-08-05 | 2023-06-23 | 中外制药株式会社 | Compositions for preventing or treating IL-8 related diseases |
WO2018029124A1 (en) | 2016-08-08 | 2018-02-15 | F. Hoffmann-La Roche Ag | Therapeutic and diagnostic methods for cancer |
JP7213549B2 (en) | 2016-08-22 | 2023-01-27 | シーエイチオー ファーマ インコーポレイテッド | Antibodies, Binding Fragments, and Methods of Use |
WO2018045379A1 (en) | 2016-09-02 | 2018-03-08 | Dana-Farber Cancer Institute, Inc. | Composition and methods of treating b cell disorders |
EP3510046A4 (en) | 2016-09-07 | 2020-05-06 | The Regents of the University of California | Antibodies to oxidation-specific epitopes |
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 |
CN109689682B (en) | 2016-09-19 | 2022-11-29 | 豪夫迈·罗氏有限公司 | Complement factor-based affinity chromatography |
LT3528838T (en) | 2016-09-23 | 2023-10-10 | F. Hoffmann-La Roche Ag | Uses of il-13 antagonists for treating atopic dermatitis |
JOP20190055A1 (en) | 2016-09-26 | 2019-03-24 | Merck Sharp & Dohme | Anti-cd27 antibodies |
CA3037961A1 (en) | 2016-09-30 | 2018-04-05 | Janssen Biotech, Inc. | Safe and effective method of treating psoriasis with anti-il23 specific antibody |
MX2019003768A (en) | 2016-10-03 | 2019-06-24 | Juno Therapeutics Inc | Hpv-specific binding molecules. |
WO2018065501A1 (en) | 2016-10-05 | 2018-04-12 | F. Hoffmann-La Roche Ag | Methods for preparing antibody drug conjugates |
MX2019003934A (en) | 2016-10-06 | 2019-07-10 | Genentech Inc | Therapeutic and diagnostic methods for cancer. |
US20200023072A1 (en) | 2016-10-11 | 2020-01-23 | Medimmune Limited | Antibody-drug conjugates with immune-mediated therapy agents |
WO2018068201A1 (en) | 2016-10-11 | 2018-04-19 | Nanjing Legend Biotech Co., Ltd. | Single-domain antibodies and variants thereof against ctla-4 |
EP3532091A2 (en) | 2016-10-29 | 2019-09-04 | H. Hoffnabb-La Roche Ag | Anti-mic antibidies and methods of use |
PL3535298T3 (en) | 2016-11-02 | 2021-12-27 | Jounce Therapeutics, Inc. | Antibodies to pd-1 and uses thereof |
MX2019005438A (en) | 2016-11-15 | 2019-08-16 | Genentech Inc | Dosing for treatment with anti-cd20/anti-cd3 bispecific antibodies. |
KR20190078648A (en) | 2016-11-16 | 2019-07-04 | 얀센 바이오테크 인코포레이티드 | Methods for treating psoriasis with anti-IL23 specific antibodies |
JOP20190100A1 (en) | 2016-11-19 | 2019-05-01 | Potenza Therapeutics Inc | Anti-gitr antigen-binding proteins and methods of use thereof |
NZ750948A (en) | 2016-11-21 | 2020-06-26 | Cureab Gmbh | Anti-gp73 antibodies and immunoconjugates |
US10759855B2 (en) | 2016-12-02 | 2020-09-01 | Rigel Pharmaceuticals, Inc. | Antigen binding molecules to TIGIT |
KR102603681B1 (en) | 2016-12-07 | 2023-11-17 | 아게누스 인코포레이티드 | Antibodies and methods of using them |
AU2017373889A1 (en) | 2016-12-07 | 2019-06-06 | Ac Immune Sa | Anti-Tau antibodies and methods of use |
CN110248959B (en) | 2016-12-07 | 2023-06-30 | 基因泰克公司 | anti-TAU antibodies and methods of use |
CN110366562A (en) | 2016-12-12 | 2019-10-22 | 豪夫迈·罗氏有限公司 | Use the method for anti-PD-L1 antibody and anti-androgen therapy cancer |
TWI829628B (en) | 2016-12-19 | 2024-01-21 | 瑞士商赫孚孟拉羅股份公司 | Combination therapy with targeted 4-1bb (cd137) agonists |
JP7247091B2 (en) | 2016-12-20 | 2023-03-28 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Combination therapy with anti-CD20/anti-CD3 bispecific antibody and 4-1BB (CD137) agonist |
JOP20190134A1 (en) | 2016-12-23 | 2019-06-02 | Potenza Therapeutics Inc | Anti-neuropilin antigen-binding proteins and methods of use thereof |
TW201829469A (en) | 2017-01-03 | 2018-08-16 | 瑞士商赫孚孟拉羅股份公司 | Bispecific antigen binding molecules comprising anti-4-1bb clone 20h4.9 |
WO2018129029A1 (en) | 2017-01-04 | 2018-07-12 | Immunogen, Inc. | Met antibodies and immunoconjugates and uses thereof |
TW201831517A (en) | 2017-01-12 | 2018-09-01 | 美商優瑞科生物技術公司 | Constructs targeting histone h3 peptide/mhc complexes and uses thereof |
US11266745B2 (en) | 2017-02-08 | 2022-03-08 | Imaginab, Inc. | Extension sequences for diabodies |
UA126574C2 (en) | 2017-02-10 | 2022-11-02 | Дженентек, Інк. | Anti-tryptase antibodies, compositions thereof, and uses thereof |
US11021535B2 (en) | 2017-02-10 | 2021-06-01 | The United States Of America As Represented By The Secretary, Department Of Health And Human Services | Neutralizing antibodies to plasmodium falciparum circumsporozoite protein and their use |
EP3589754B1 (en) | 2017-03-01 | 2023-06-28 | F. Hoffmann-La Roche AG | Diagnostic and therapeutic methods for cancer |
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 |
SG10201911225WA (en) | 2017-03-28 | 2020-01-30 | Genentech Inc | Methods of treating neurodegenerative diseases |
WO2018178076A1 (en) | 2017-03-29 | 2018-10-04 | F. Hoffmann-La Roche Ag | Bispecific antigen binding molecule for a costimulatory tnf receptor |
WO2018178055A1 (en) | 2017-03-29 | 2018-10-04 | F. Hoffmann-La Roche Ag | Bispecific antigen binding molecule for a costimulatory tnf receptor |
JOP20190203A1 (en) | 2017-03-30 | 2019-09-03 | Potenza Therapeutics Inc | Anti-tigit antigen-binding proteins and methods of use thereof |
CA3055132A1 (en) | 2017-04-03 | 2018-10-11 | F. Hoffmann-La Roche Ag | Antibodies binding to steap-1 |
BR112019017753A2 (en) | 2017-04-04 | 2020-04-07 | Hoffmann La Roche | bispecific molecule, polynucleotide, vector, cell, methods for the production of a molecule and for the treatment of an individual, composition and use of the bispecific molecule |
SI3606954T1 (en) | 2017-04-05 | 2022-10-28 | F. Hoffmann - La Roche Ag | Anti-lag3 antibodies |
KR102408873B1 (en) | 2017-04-05 | 2022-06-15 | 에프. 호프만-라 로슈 아게 | Bispecific antibodies specifically binding to pd1 and lag3 |
MX2019012192A (en) | 2017-04-14 | 2020-01-21 | Genentech Inc | Diagnostic and therapeutic methods for cancer. |
EP3624820A1 (en) | 2017-04-21 | 2020-03-25 | H. Hoffnabb-La Roche Ag | Use of klk5 antagonists for treatment of a disease |
AU2018258049A1 (en) | 2017-04-26 | 2019-12-12 | Eureka Therapeutics, Inc. | Constructs specifically recognizing glypican 3 and uses thereof |
US20220135670A1 (en) | 2017-04-27 | 2022-05-05 | Tesaro, Inc. | Antibody agents directed against lymphocyte activation gene-3 (lag-3) and uses thereof |
US11203638B2 (en) | 2017-05-05 | 2021-12-21 | Allakos Inc. | Methods and compositions for treating perennial allergic conjunctivitis and keratoconjunctivitis |
JP7299842B2 (en) | 2017-05-16 | 2023-06-28 | ファイヴ プライム セラピューティクス インク | Anti-FGFR2 Antibodies in Combination with Chemotherapeutic Agents in Cancer Treatment |
EP3630829A1 (en) | 2017-06-02 | 2020-04-08 | H. Hoffnabb-La Roche Ag | Type ii anti-cd20 antibody and anti-cd20/cd3 bispecific antibody for treatment of cancer |
US11634488B2 (en) | 2017-07-10 | 2023-04-25 | International—Drug—Development—Biotech | Treatment of B cell malignancies using afucosylated pro-apoptotic anti-CD19 antibodies in combination with anti CD20 antibodies or chemotherapeutics |
KR20200093518A (en) | 2017-07-21 | 2020-08-05 | 제넨테크, 인크. | Methods of treatment and diagnosis for cancer |
US10961318B2 (en) | 2017-07-26 | 2021-03-30 | Forty Seven, Inc. | Anti-SIRP-α antibodies and related methods |
BR112020003533A2 (en) | 2017-08-25 | 2020-11-17 | Five Prime Therapeutics, Inc. | b7-h4 antibodies and methods of using them |
EP3684413A1 (en) | 2017-09-20 | 2020-07-29 | Chugai Seiyaku Kabushiki Kaisha | Dosage regimen for combination therapy using pd-1 axis binding antagonists and gpc3 targeting agent |
TW201922780A (en) | 2017-09-25 | 2019-06-16 | 美商健生生物科技公司 | Safe and effective method of treating Lupus with anti-IL12/IL23 antibody |
CA3080546A1 (en) | 2017-10-03 | 2019-04-11 | Juno Therapeutics, Inc. | Hpv-specific binding molecules |
WO2019075090A1 (en) | 2017-10-10 | 2019-04-18 | Tilos Therapeutics, Inc. | Anti-lap antibodies and uses thereof |
CA3078974A1 (en) | 2017-10-12 | 2019-04-18 | Immunowake Inc. | Vegfr-antibody light chain fusion protein |
EP3703746A1 (en) | 2017-11-01 | 2020-09-09 | F. Hoffmann-La Roche AG | Novel tnf family ligand trimer-containing antigen binding molecules |
JP2021501162A (en) | 2017-11-01 | 2021-01-14 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | Combination therapy with targeted OX40 agonist |
SG11202003501XA (en) | 2017-11-01 | 2020-05-28 | Juno Therapeutics Inc | Antibodies and chimeric antigen receptors specific for b-cell maturation antigen |
WO2019086500A2 (en) | 2017-11-01 | 2019-05-09 | F. Hoffmann-La Roche Ag | Bispecific 2+1 contorsbodies |
MX2020004567A (en) | 2017-11-06 | 2020-08-13 | Genentech Inc | Diagnostic and therapeutic methods for cancer. |
MX2020005662A (en) | 2017-12-01 | 2020-08-20 | Pfizer | Anti-cxcr5 antibodies and compositions and uses thereof. |
AU2018380979B2 (en) | 2017-12-08 | 2023-07-20 | Argenx Bvba | Use of FcRn antagonists for treatment of generalized myasthenia gravis |
KR20200110745A (en) | 2017-12-15 | 2020-09-25 | 주노 쎄러퓨티크스 인코퍼레이티드 | Anti-CCT5 binding molecule and method of use thereof |
TWI805665B (en) | 2017-12-21 | 2023-06-21 | 瑞士商赫孚孟拉羅股份公司 | Antibodies binding to hla-a2/wt1 |
EP3502140A1 (en) | 2017-12-21 | 2019-06-26 | F. Hoffmann-La Roche AG | Combination therapy of tumor targeted icos agonists with t-cell bispecific molecules |
US20190211098A1 (en) | 2017-12-22 | 2019-07-11 | Genentech, Inc. | Use of pilra binding agents for treatment of a disease |
WO2019126514A2 (en) | 2017-12-22 | 2019-06-27 | Jounce Therapeutics, Inc. | Antibodies for lilrb2 |
WO2019129221A1 (en) | 2017-12-28 | 2019-07-04 | Nanjing Legend Biotech Co., Ltd. | Single-domain antibodies and variants thereof against tigit |
KR20200104886A (en) | 2017-12-28 | 2020-09-04 | 난징 레전드 바이오테크 씨오., 엘티디. | Antibodies and variants against PD-L1 |
WO2019129679A1 (en) | 2017-12-29 | 2019-07-04 | F. Hoffmann-La Roche Ag | Method for improving vegf-receptor blocking selectivity of an anti-vegf antibody |
WO2019136029A1 (en) | 2018-01-02 | 2019-07-11 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Neutralizing antibodies to ebola virus glycoprotein and their use |
CN111886247A (en) | 2018-01-05 | 2020-11-03 | Ac免疫有限公司 | Misfolded TDP-43 binding molecules |
EP3508499A1 (en) | 2018-01-08 | 2019-07-10 | iOmx Therapeutics AG | Antibodies targeting, and other modulators of, an immunoglobulin gene associated with resistance against anti-tumour immune responses, 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 |
JP7345479B2 (en) | 2018-01-26 | 2023-09-15 | ジェネンテック, インコーポレイテッド | Composition and method of use |
JP7349995B2 (en) | 2018-01-26 | 2023-09-25 | ジェネンテック, インコーポレイテッド | IL-22 Fc fusion protein and method of use |
CN116041516A (en) | 2018-02-01 | 2023-05-02 | 信达生物制药(苏州)有限公司 | Fully human anti-B Cell Maturation Antigen (BCMA) single-chain antibody and application thereof |
AR115360A1 (en) | 2018-02-08 | 2021-01-13 | Genentech Inc | ANTIGEN BINDING MOLECULES AND METHODS OF USE |
TWI829667B (en) | 2018-02-09 | 2024-01-21 | 瑞士商赫孚孟拉羅股份公司 | Antibodies binding to gprc5d |
MA51741A (en) | 2018-02-09 | 2021-05-19 | Hoffmann La Roche | THERAPEUTIC AND DIAGNOSIS PROCEDURES FOR INFLAMMATORY DISEASES MEDIATED BY MASTOCYTES |
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 |
KR20200123169A (en) | 2018-02-21 | 2020-10-28 | 파이브 프라임 테라퓨틱스, 인크. | B7-H4 antibody dosing regimen |
MX2020008736A (en) | 2018-02-21 | 2020-12-07 | Five Prime Therapeutics Inc | B7-h4 antibody formulations. |
WO2019165122A1 (en) | 2018-02-21 | 2019-08-29 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Neutralizing antibodies to hiv-1 env and their use |
EP3755364A1 (en) | 2018-02-21 | 2020-12-30 | F. Hoffmann-La Roche AG | Dosing for treatment with il-22 fc fusion proteins |
CA3092108A1 (en) | 2018-02-26 | 2019-08-29 | Genentech, Inc. | Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies |
AU2019228600A1 (en) | 2018-03-02 | 2020-09-24 | Five Prime Therapeutics, Inc. | B7-H4 antibodies and methods of use thereof |
KR20200129125A (en) | 2018-03-05 | 2020-11-17 | 얀센 바이오테크 인코포레이티드 | How to treat Crohn's disease with anti-IL23 specific antibodies |
TW202003561A (en) | 2018-03-13 | 2020-01-16 | 瑞士商赫孚孟拉羅股份公司 | Combination therapy with targeted 4-1BB (CD137) agonists |
AU2019236372A1 (en) | 2018-03-13 | 2020-07-30 | F. Hoffmann-La Roche Ag | Therapeutic combination of 4-1 BB agonists with anti-CD20 antibodies |
CN112166123B (en) | 2018-03-14 | 2022-09-30 | 北京轩义医药科技有限公司 | Anti-claudin 18.2 antibodies |
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 |
TW201945393A (en) | 2018-03-21 | 2019-12-01 | 美商戊瑞治療有限公司 | Antibodies binding to VISTA at acidic pH |
JP2021519073A (en) | 2018-03-29 | 2021-08-10 | ジェネンテック, インコーポレイテッド | Regulation of lactogenic activity in mammalian cells |
TW202011029A (en) | 2018-04-04 | 2020-03-16 | 美商建南德克公司 | Methods for detecting and quantifying FGF21 |
CA3095027A1 (en) | 2018-04-05 | 2019-10-10 | Juno Therapeutics, Inc. | T cell receptors and engineered cells expressing same |
EP3774900A1 (en) | 2018-04-13 | 2021-02-17 | F. Hoffmann-La Roche AG | Her2-targeting antigen binding molecules comprising 4-1bbl |
US20190345245A1 (en) | 2018-05-11 | 2019-11-14 | Janssen Biotech, Inc. | Methods of Treating Crohn's Disease with Anti-IL23 Specific Antibody |
AU2019271148B2 (en) | 2018-05-14 | 2023-07-06 | Werewolf Therapeutics, Inc. | Activatable interleukin-2 polypeptides and methods of use thereof |
BR112020023118A2 (en) | 2018-05-14 | 2021-04-13 | Werewolf Therapeutics, Inc. | ACTIVE ACTIVABLE INTERLEUKIN POLYPEPTIDS 12 AND METHODS OF USE OF THESE |
WO2019224275A1 (en) | 2018-05-23 | 2019-11-28 | Adc Therapeutics Sa | Molecular adjuvant |
WO2019234576A1 (en) | 2018-06-03 | 2019-12-12 | Lamkap Bio Beta Ltd. | Bispecific antibodies against ceacam5 and cd47 |
JP7372237B2 (en) | 2018-06-04 | 2023-10-31 | 中外製薬株式会社 | Antigen-binding molecules with altered half-lives in the cytoplasm |
BR112020026384A2 (en) | 2018-06-23 | 2021-03-30 | Genentech, Inc. | METHODS FOR TREATING AN INDIVIDUAL WITH LUNG CANCER AND FOR TREATING AN INDIVIDUAL WITH SMALL CELL LUNG CANCER, KITS, ANTIBODY ANTI-PD-L1 AND COMPOSITION |
CN112424228A (en) | 2018-07-04 | 2021-02-26 | 豪夫迈·罗氏有限公司 | Novel bispecific agonistic 4-1BB antigen-binding molecules |
SG11202100096XA (en) | 2018-07-09 | 2021-02-25 | Five Prime Therapeutics Inc | Antibodies binding to ilt4 |
WO2020014306A1 (en) | 2018-07-10 | 2020-01-16 | Immunogen, Inc. | Met antibodies and immunoconjugates and uses thereof |
CA3104536A1 (en) | 2018-07-11 | 2020-01-16 | Bristol-Myers Squibb Company | Antibodies binding to vista at acidic ph |
WO2020016838A2 (en) | 2018-07-18 | 2020-01-23 | Janssen Biotech, Inc. | Sustained response predictors after treatment with anti-il23 specific antibody |
US20200171146A1 (en) | 2018-07-18 | 2020-06-04 | Genentech, Inc. | Methods of treating lung cancer with a pd-1 axis binding antagonist, an antimetabolite, and a platinum agent |
EP3823673A4 (en) | 2018-07-20 | 2022-05-11 | Surface Oncology, Inc. | Anti-cd112r compositions and methods |
US20220195045A1 (en) | 2018-08-03 | 2022-06-23 | Chugai Seiyaku Kabushiki Kaisha | Antigen-binding molecule containing two antigen-binding domains that are 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 |
BR112021002037A2 (en) | 2018-08-10 | 2021-05-04 | Chugai Seiyaku Kabushiki Kaisha | anti-cd137 antigen binding molecule and its use |
BR112021003156A2 (en) | 2018-08-23 | 2021-05-11 | Seagen, Inc. | composition, antibody that binds to human tigi, pharmaceutical formulation, isolated polynucleotide, vector, host cell, methods for producing an antibody that binds to human tigi and fucosylated antibodies that bind to tigi, and for the treatment of cancer , and, kit. |
GB201814281D0 (en) | 2018-09-03 | 2018-10-17 | Femtogenix Ltd | Cytotoxic agents |
EP3853611A1 (en) | 2018-09-19 | 2021-07-28 | F. Hoffmann-La Roche AG | Therapeutic and diagnostic methods for bladder cancer |
EP3857230B1 (en) | 2018-09-21 | 2023-06-07 | F. Hoffmann-La Roche AG | Diagnostic methods for triple-negative breast cancer |
CN113166236A (en) | 2018-09-21 | 2021-07-23 | 北卡罗来纳大学查佩尔希尔分校 | Synthetic adhesive with limited penetration through mucus |
SI3883606T1 (en) | 2018-09-24 | 2023-10-30 | Janssen Biotech, Inc. | Safe and effective method of treating ulcerative colitis with anti-il12/il23 antibody |
JP2022502088A (en) | 2018-09-27 | 2022-01-11 | エクシリオ デベロップメント, インコーポレイテッド | Masked cytokine polypeptide |
CN112654641A (en) | 2018-10-01 | 2021-04-13 | 豪夫迈·罗氏有限公司 | Bispecific antigen binding molecules with trivalent binding to CD40 |
US11242396B2 (en) | 2018-10-01 | 2022-02-08 | Hoffmann-La Roche Inc. | Bispecific antigen binding molecules comprising anti-FAP clone 212 |
EP3632929A1 (en) | 2018-10-02 | 2020-04-08 | Ospedale San Raffaele S.r.l. | Antibodies and uses thereof |
KR20210074286A (en) | 2018-10-05 | 2021-06-21 | 파이브 프라임 테라퓨틱스, 인크. | Anti-FGFR2 antibody formulation |
WO2020076969A2 (en) | 2018-10-10 | 2020-04-16 | Tilos Therapeutics, Inc. | Anti-lap antibody variants and uses thereof |
MX2021004226A (en) | 2018-10-15 | 2021-07-15 | Five Prime Therapeutics Inc | Combination therapy for cancer. |
WO2020081493A1 (en) | 2018-10-16 | 2020-04-23 | Molecular Templates, Inc. | Pd-l1 binding proteins |
KR20210079311A (en) | 2018-10-18 | 2021-06-29 | 제넨테크, 인크. | Diagnosis and treatment methods for sarcoma renal cancer |
US20210395390A1 (en) | 2018-10-31 | 2021-12-23 | Bayer Aktiengesellschaft | Reversal agents for neutralizing the therapeutic activity of anti-fxia antibodies |
US20210403597A1 (en) | 2018-11-16 | 2021-12-30 | Memorial Sloan Kettering Cancer Center | Antibodies to mucin-16 and methods of use thereof |
US11548941B2 (en) | 2018-11-20 | 2023-01-10 | Janssen Biotech, Inc. | Safe and effective method of treating psoriasis with anti-IL-23 specific antibody |
JP2022511502A (en) | 2018-12-05 | 2022-01-31 | ジェネンテック, インコーポレイテッド | Diagnostic Methods and Diagnostic Compositions for Cancer Immunotherapy |
EP3892299A4 (en) | 2018-12-07 | 2022-11-30 | ONO Pharmaceutical Co., Ltd. | Immunosuppressant |
WO2020123275A1 (en) | 2018-12-10 | 2020-06-18 | Genentech, Inc. | Photocrosslinking peptides for site specific conjugation to fc-containing proteins |
EP3897722A4 (en) | 2018-12-18 | 2022-09-14 | Janssen Biotech, Inc. | Safe and effective method of treating lupus with anti-il12/il23 antibody |
US20220089694A1 (en) | 2018-12-20 | 2022-03-24 | The U.S.A., As Represented By The Secretary, Department Of Health And Human Services | Ebola virus glycoprotein-specific monoclonal antibodies and uses thereof |
AR117453A1 (en) | 2018-12-20 | 2021-08-04 | Genentech Inc | CF OF MODIFIED ANTIBODIES AND METHODS TO USE THEM |
MA54514A (en) | 2018-12-21 | 2022-03-30 | Hoffmann La Roche | ANTIBODIES BINDING TO CD3 |
EP3898682A1 (en) | 2018-12-21 | 2021-10-27 | F. Hoffmann-La Roche AG | Tumor-targeted agonistic cd28 antigen binding molecules |
MX2021007421A (en) | 2018-12-21 | 2021-08-05 | Hoffmann La Roche | Antibody that binds to vegf and il-1beta and methods of use. |
JP2022513495A (en) | 2018-12-21 | 2022-02-08 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Tumor targeting superagonist CD28 antigen binding molecule |
EP3902832A2 (en) | 2018-12-26 | 2021-11-03 | Xilio Development, Inc. | Anti-ctla4 antibodies and methods of use thereof |
CN113272327A (en) | 2018-12-30 | 2021-08-17 | 豪夫迈·罗氏有限公司 | Anti-rabbit CD19 antibodies and methods of use thereof |
TW202043272A (en) | 2019-01-14 | 2020-12-01 | 美商建南德克公司 | Methods of treating cancer with a pd-1 axis binding antagonist and an rna vaccine |
EP3911675A1 (en) | 2019-01-17 | 2021-11-24 | Bayer Aktiengesellschaft | Methods to determine whether a subject is suitable of being treated with an agonist of soluble guanylyl cyclase (sgc) |
CN113329770A (en) | 2019-01-24 | 2021-08-31 | 中外制药株式会社 | Novel cancer antigen and antibody against said antigen |
GB201901197D0 (en) | 2019-01-29 | 2019-03-20 | Femtogenix Ltd | G-A Crosslinking cytotoxic agents |
KR20210122272A (en) | 2019-01-29 | 2021-10-08 | 주노 쎄러퓨티크스 인코퍼레이티드 | Antibodies and chimeric antigen receptors specific for receptor tyrosine kinase-like orphan receptor 1 (ROR1) |
CA3130695A1 (en) | 2019-02-27 | 2020-09-03 | Genentech, Inc. | Dosing for treatment with anti-tigit and anti-cd20 or anti-cd38 antibodies |
WO2020185535A1 (en) | 2019-03-08 | 2020-09-17 | Genentech, Inc. | Methods for detecting and quantifying membrane-associated proteins on extracellular vesicles |
JP2022524074A (en) | 2019-03-14 | 2022-04-27 | ジェネンテック, インコーポレイテッド | Treatment of cancer with HER2xCD3 bispecific antibodies in combination with anti-HER2 MAB |
JP2022525145A (en) | 2019-03-14 | 2022-05-11 | ヤンセン バイオテツク,インコーポレーテツド | A production method for producing an anti-IL12 / IL23 antibody composition. |
EA202192459A1 (en) | 2019-03-18 | 2021-11-25 | Янссен Байотек, Инк. | METHOD FOR TREATMENT OF PSORIASIS WITH ANTIBODY TO IL12 / IL23 IN CHILDREN |
GB2589049C (en) | 2019-04-11 | 2024-02-21 | argenx BV | Anti-IgE antibodies |
JP7301155B2 (en) | 2019-04-12 | 2023-06-30 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Bispecific antigen-binding molecules containing lipocalin muteins |
MX2021012692A (en) | 2019-04-19 | 2021-11-12 | Genentech Inc | Anti-mertk antibodies and their methods of use. |
SG11202111345PA (en) | 2019-04-19 | 2021-11-29 | Chugai Pharmaceutical Co Ltd | Chimeric receptor that recognizes engineered site in antibody |
US20220227853A1 (en) | 2019-05-03 | 2022-07-21 | The United States Of America,As Represented By The Secretary,Department Of Health And Human Services | Neutralizing antibodies to plasmodium falciparum circumsporozoite protein and their use |
CN114269376A (en) | 2019-05-03 | 2022-04-01 | 豪夫迈·罗氏有限公司 | Methods of treating cancer with anti-PD-L1 antibodies |
EP3969035A4 (en) | 2019-05-14 | 2023-06-21 | Werewolf Therapeutics, Inc. | Separation moieties and methods and use thereof |
CA3138045C (en) | 2019-05-14 | 2024-02-20 | Genentech, Inc. | Methods of using anti-cd79b immunoconjugates to treat follicular lymphoma |
US20230085439A1 (en) | 2019-05-21 | 2023-03-16 | University Of Georgia Research Foundation, Inc. | Antibodies that bind human metapneumovirus fusion protein and their use |
KR20220012883A (en) | 2019-05-23 | 2022-02-04 | 얀센 바이오테크 인코포레이티드 | A method of treating inflammatory bowel disease with a combination therapy of IL-23 and an antibody against TNF alpha |
AU2020278907A1 (en) | 2019-05-23 | 2022-01-20 | Ac Immune Sa | Anti-TDP-43 binding molecules and uses thereof |
CA3138072A1 (en) | 2019-06-07 | 2020-12-10 | Argenx Bvba | Pharmaceutical formulations of fcrn inhibitors suitable for subcutaneous administration |
AU2020304813A1 (en) | 2019-06-26 | 2022-01-06 | F. Hoffmann-La Roche Ag | Fusion of an antibody binding CEA and 4-1BBL |
JP7354306B2 (en) | 2019-06-27 | 2023-10-02 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Novel ICOS antibodies and tumor-targeting antigen-binding molecules containing them |
EA202290208A1 (en) | 2019-07-02 | 2022-03-25 | Дзе Юнайтед Стейтс Оф Эмерика, Эз Репрезентед Бай Дзе Секретэри, Дипартмент Оф Хелт Энд Хьюман Сервисиз | MONOCLONAL ANTIBODIES THAT BIND EGFRvIII AND THEIR APPLICATIONS |
CA3146023A1 (en) | 2019-07-05 | 2021-01-14 | Iomx Therapeutics Ag | Antibodies binding igc2 of igsf11 (vsig3) and uses thereof |
AR119382A1 (en) | 2019-07-12 | 2021-12-15 | Hoffmann La Roche | PRE-TARGETING ANTIBODIES AND METHODS OF USE |
JPWO2021010326A1 (en) | 2019-07-12 | 2021-01-21 | ||
AR119393A1 (en) | 2019-07-15 | 2021-12-15 | Hoffmann La Roche | ANTIBODIES THAT BIND NKG2D |
KR20220038415A (en) | 2019-07-26 | 2022-03-28 | 벤더르빌트 유니버시티 | Human monoclonal antibody to enterovirus D68 |
SG11202112491WA (en) | 2019-07-31 | 2021-12-30 | Hoffmann La Roche | Antibodies binding to gprc5d |
EP4004045A1 (en) | 2019-07-31 | 2022-06-01 | F. Hoffmann-La Roche AG | Antibodies binding to gprc5d |
WO2021024209A1 (en) | 2019-08-06 | 2021-02-11 | Aprinoia Therapeutics Inc. | Antibodies that bind to pathological tau species and uses thereof |
TW202118512A (en) | 2019-09-12 | 2021-05-16 | 美商建南德克公司 | Compositions and methods of treating lupus nephritis |
EP4031579A2 (en) | 2019-09-18 | 2022-07-27 | F. Hoffmann-La Roche AG | Anti-klk7 antibodies, anti-klk5 antibodies, multispecific anti-klk5/klk7 antibodies, and methods of use |
EP4031575A1 (en) | 2019-09-19 | 2022-07-27 | Bristol-Myers Squibb Company | Antibodies binding to vista at acidic ph |
EP4031580A1 (en) | 2019-09-20 | 2022-07-27 | F. Hoffmann-La Roche AG | Dosing for anti-tryptase antibodies |
KR20220070237A (en) | 2019-09-27 | 2022-05-30 | 제넨테크, 인크. | Dosing for treatment with anti-TIGIT and anti-PD-L1 antagonist antibodies |
AU2020365836A1 (en) | 2019-10-18 | 2022-04-28 | F. Hoffmann-La Roche Ag | Methods of using anti-CD79b immunoconjugates to treat diffuse large B-cell lymphoma |
EP4055388A1 (en) | 2019-11-06 | 2022-09-14 | Genentech, Inc. | Diagnostic and therapeutic methods for treatment of hematologic cancers |
BR112022008629A2 (en) | 2019-11-15 | 2022-07-19 | Enthera S R L | ISOLATED ANTIBODY OR ANTIGEN BINDING FRAGMENT THEREOF, ISOLATED POLYNUCLEOTIDE, VECTOR, ISOLATED CELL, PHARMACEUTICAL COMPOSITION, USE THEREOF AND METHOD FOR INHIBITING IGFBP3 BINDING TO THE TMEM219 RECEPTOR |
EP3822288A1 (en) | 2019-11-18 | 2021-05-19 | Deutsches Krebsforschungszentrum, Stiftung des öffentlichen Rechts | Antibodies targeting, and other modulators of, the cd276 antigen, and uses thereof |
US20230039165A1 (en) | 2019-11-21 | 2023-02-09 | Enthera S.R.L. | Igfbp3 antibodies and therapeutic uses thereof |
EP3831849A1 (en) | 2019-12-02 | 2021-06-09 | LamKap Bio beta AG | Bispecific antibodies against ceacam5 and cd47 |
CN115335399A (en) | 2019-12-06 | 2022-11-11 | 朱诺治疗学股份有限公司 | Anti-idiotype antibodies directed against GPRC5D target binding domains and related compositions and methods |
BR112022010627A2 (en) | 2019-12-06 | 2022-08-16 | Juno Therapeutics Inc | ANTI-IDIOTYPIC ANTIBODIES TO BINDING DOMAINS TARGETED BY BCMA AND RELATED COMPOSITIONS AND METHODS |
WO2021119505A1 (en) | 2019-12-13 | 2021-06-17 | Genentech, Inc. | Anti-ly6g6d antibodies and methods of use |
US20210230278A1 (en) | 2019-12-18 | 2021-07-29 | Hoffmann-La Roche Inc. | Antibodies binding to HLA-A2/MAGE-A4 |
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 |
US20230058982A1 (en) | 2019-12-27 | 2023-02-23 | Chugai Seiyaku Kabushiki Kaisha | Anti-ctla-4 antibody and use thereof |
JP2023509195A (en) | 2020-01-08 | 2023-03-07 | アルジェニクス ビーブイ | How to treat pemphigus |
MX2022008214A (en) | 2020-01-09 | 2022-08-08 | Hoffmann La Roche | New 4-1bbl trimer-containing antigen binding molecules. |
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 |
WO2022050954A1 (en) | 2020-09-04 | 2022-03-10 | Genentech, Inc. | Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies |
WO2021194481A1 (en) | 2020-03-24 | 2021-09-30 | Genentech, Inc. | Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies |
CA3164559A1 (en) | 2020-01-31 | 2021-08-05 | Lars Mueller | Methods of inducing neoepitope-specific t cells with a pd-1 axis binding antagonist and an rna vaccine |
WO2021155295A1 (en) | 2020-01-31 | 2021-08-05 | The Cleveland Clinic Foundation | Anti-müllerian hormone receptor 2 antibodies and methods of use |
AU2021220847A1 (en) | 2020-02-11 | 2022-09-01 | Vanderbilt University | Human monoclonal antibodies to severe acute respiratory syndrome coronavirus 2 (SARS-CoV- 2) |
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 |
CA3169451A1 (en) | 2020-02-14 | 2021-08-19 | Jounce Therapeutics, Inc. | Antibodies and fusion proteins that bind to ccr8 and uses thereof |
WO2021168292A1 (en) | 2020-02-20 | 2021-08-26 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Epstein-barr virus monoclonal antibodies and uses thereof |
EP3868396A1 (en) | 2020-02-20 | 2021-08-25 | Enthera S.R.L. | Inhibitors and uses thereof |
JP2023516941A (en) | 2020-02-28 | 2023-04-21 | 上海復宏漢霖生物技術股▲フン▼有限公司 | Anti-CD137 constructs, multispecific antibodies and uses thereof |
JP2023516945A (en) | 2020-02-28 | 2023-04-21 | 上海復宏漢霖生物技術股▲フン▼有限公司 | Anti-CD137 constructs and uses thereof |
CN115315446A (en) | 2020-03-06 | 2022-11-08 | Go医疗股份有限公司 | anti-sugar-CD 44 antibodies and uses thereof |
CR20220461A (en) | 2020-03-13 | 2022-10-21 | 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 |
CN115867577A (en) | 2020-03-23 | 2023-03-28 | 基因泰克公司 | Biomarkers for predicting response to IL-6 antagonists in COVID-19 pneumonia |
EP4107184A1 (en) | 2020-03-23 | 2022-12-28 | Genentech, Inc. | Method for treating pneumonia, including covid-19 pneumonia, with an il6 antagonist |
US20230174656A1 (en) | 2020-03-23 | 2023-06-08 | Genentech, Inc. | Tocilizumab and remdesivir combination therapy for covid-19 pneumonia |
PE20230414A1 (en) | 2020-03-24 | 2023-03-07 | Genentech Inc | TIE2 FIXING AGENTS AND METHODS OF USE |
MX2022011892A (en) | 2020-03-26 | 2022-10-18 | Univ Vanderbilt | Human monoclonal antibodies to severe acute respiratory syndrome coronavirus 2 (sars-cov-2). |
WO2021195385A1 (en) | 2020-03-26 | 2021-09-30 | Vanderbilt University | HUMAN MONOCLONAL ANTIBODIES TO SEVERE ACUTE RESPIRATORY SYNDROME CORONAVIRUS 2 (SARS-GoV-2) |
TW202202620A (en) | 2020-03-26 | 2022-01-16 | 美商建南德克公司 | Modified mammalian cells |
AR121706A1 (en) | 2020-04-01 | 2022-06-29 | Hoffmann La Roche | OX40 AND FAP-TARGETED BSPECIFIC ANTIGEN-BINDING MOLECULES |
WO2021202959A1 (en) | 2020-04-03 | 2021-10-07 | Genentech, Inc. | Therapeutic and diagnostic methods for cancer |
EP4132971A1 (en) | 2020-04-09 | 2023-02-15 | Merck Sharp & Dohme LLC | 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 |
MX2022013198A (en) | 2020-04-24 | 2022-11-14 | Genentech Inc | Methods of using anti-cd79b immunoconjugates. |
KR20230004520A (en) | 2020-04-27 | 2023-01-06 | 더 리젠츠 오브 더 유니버시티 오브 캘리포니아 | Isotype-independent antibodies to lipoproteins (a) |
JP2023523450A (en) | 2020-04-28 | 2023-06-05 | ジェネンテック, インコーポレイテッド | Methods and compositions for non-small cell lung cancer immunotherapy |
CN116963782A (en) | 2020-05-03 | 2023-10-27 | 联宁(苏州)生物制药有限公司 | Antibody drug conjugates comprising anti-TROP-2 antibodies |
AU2021275361A1 (en) | 2020-05-17 | 2023-01-19 | Astrazeneca Uk Limited | SARS-CoV-2 antibodies and methods of selecting and using the same |
US20230220057A1 (en) | 2020-05-27 | 2023-07-13 | Staidson (Beijing) Biopharmaceuticals Co., Ltd. | Antibodies specifically recognizing nerve growth factor and uses thereof |
MX2022015376A (en) | 2020-06-02 | 2023-04-14 | 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 |
CN115697489A (en) | 2020-06-08 | 2023-02-03 | 豪夫迈·罗氏有限公司 | anti-HBV antibodies and methods of use thereof |
WO2021252977A1 (en) | 2020-06-12 | 2021-12-16 | Genentech, Inc. | Methods and compositions for cancer immunotherapy |
CA3181820A1 (en) | 2020-06-16 | 2021-12-23 | Genentech, Inc. | Methods and compositions for treating triple-negative breast cancer |
US20210395366A1 (en) | 2020-06-18 | 2021-12-23 | Genentech, Inc. | Treatment with anti-tigit antibodies and pd-1 axis binding antagonists |
BR112022025856A2 (en) | 2020-06-19 | 2023-01-10 | Hoffmann La Roche | ANTIBODY THAT BINDS CD3 AND CD19, POLYNUCLEOTIDE ISOLATED, HOST CELL, METHOD OF PRODUCING AN ANTIBODY THAT BINDS CD3 AND CD19, PHARMACEUTICAL COMPOSITION, USE OF THE ANTIBODY, METHOD FOR TREATING A DISEASE IN A SUBJECT AND INVENTION |
CR20220639A (en) | 2020-06-19 | 2023-02-17 | Hoffmann La Roche | Antibodies binding to cd3 and folr1 |
WO2021255146A1 (en) | 2020-06-19 | 2021-12-23 | F. Hoffmann-La Roche Ag | Antibodies binding to cd3 and cea |
PE20230835A1 (en) | 2020-06-19 | 2023-05-19 | Hoffmann La Roche | ANTIBODIES THAT BIND CD3 |
EP4168448A1 (en) | 2020-06-23 | 2023-04-26 | F. Hoffmann-La Roche AG | Agonistic cd28 antigen binding molecules targeting her2 |
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 |
JP2023531067A (en) | 2020-06-25 | 2023-07-20 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Anti-CD3/Anti-CD28 Bispecific Antigen Binding Molecules |
WO2022008027A1 (en) | 2020-07-06 | 2022-01-13 | Iomx Therapeutics Ag | Antibodies binding igv of igsf11 (vsig3) and uses thereof |
CR20230076A (en) | 2020-07-10 | 2023-03-13 | 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 |
MX2023001083A (en) | 2020-07-29 | 2023-04-10 | 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 |
KR20230095918A (en) | 2020-08-05 | 2023-06-29 | 주노 쎄러퓨티크스 인코퍼레이티드 | Anti-idiotype antibodies to the ROR1-target binding domain and related compositions and methods |
BR112023002123A2 (en) | 2020-08-07 | 2023-03-07 | Genentech Inc | FC FUSION PROTEIN, ISOLATED NUCLEIC ACIDS, METHOD OF PRODUCING FC FUSION PROTEIN, PHARMACEUTICAL FORMULATION, METHODS FOR EXPANDING THE NUMBER OF DENDRITIC CELLS (DCS) IN AN INDIVIDUAL AND FOR TREATMENT OF CANCER, EFFECTOR-FREE PROTEIN AND ANTI-FCRP |
JP2023537078A (en) | 2020-08-10 | 2023-08-30 | アストラゼネカ・ユーケイ・リミテッド | SARS-CoV-2 Antibodies for Treating and Preventing COVID-19 |
JP2023537761A (en) | 2020-08-14 | 2023-09-05 | エイシー イミューン ソシエテ アノニム | Humanized anti-TDP-43 binding molecules and uses thereof |
WO2022043517A2 (en) | 2020-08-27 | 2022-03-03 | Cureab Gmbh | Anti-golph2 antibodies for macrophage and dendritic cell differentiation |
TW202227625A (en) | 2020-08-28 | 2022-07-16 | 美商建南德克公司 | Crispr/cas9 multiplex knockout of host cell proteins |
PE20230986A1 (en) | 2020-09-04 | 2023-06-21 | Hoffmann La Roche | ANTIBODY THAT BINDS VEGF-A AND ANG2, AND METHODS OF USE |
US20240010750A1 (en) | 2020-09-15 | 2024-01-11 | Bayer Aktiengesellschaft | Novel anti-a2ap antibodies and uses thereof |
US20230357418A1 (en) | 2020-09-17 | 2023-11-09 | Genentech, Inc. | Results of empacta: a randomized, double-blind, placebo-controlled, multicenter study to evaluate the efficacy and safety of tocilizumab in hospitalized patients with covid-19 pneumonia |
EP4225443A1 (en) | 2020-10-05 | 2023-08-16 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
US20230372528A1 (en) | 2020-10-16 | 2023-11-23 | University Of Georgia Research Foundation, Inc. | Glycoconjugates |
WO2022084210A1 (en) | 2020-10-20 | 2022-04-28 | F. Hoffmann-La Roche Ag | Combination therapy of pd-1 axis binding antagonists and lrrk2 inhitibors |
EP4232475A1 (en) | 2020-10-20 | 2023-08-30 | Kantonsspital St. Gallen | Antibodies or antigen-binding fragments specifically binding to gremlin-1 and uses thereof |
AR123855A1 (en) | 2020-10-20 | 2023-01-18 | Genentech Inc | PEG-CONJUGATED ANTI-MERTK ANTIBODIES AND METHODS OF USE |
WO2022093981A1 (en) | 2020-10-28 | 2022-05-05 | Genentech, Inc. | Combination therapy comprising ptpn22 inhibitors and pd-l1 binding antagonists |
EP4240762A1 (en) | 2020-11-03 | 2023-09-13 | Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts | Target-cell restricted, costimulatory, bispecific and bivalent anti-cd28 antibodies |
JP7402381B2 (en) | 2020-11-04 | 2023-12-20 | ジェネンテック, インコーポレイテッド | Administration for treatment with anti-CD20/anti-CD3 bispecific antibodies |
TW202225191A (en) | 2020-11-04 | 2022-07-01 | 美商建南德克公司 | Subcutaneous dosing of anti-cd20/anti-cd3 bispecific antibodies |
MX2023005131A (en) | 2020-11-04 | 2023-05-25 | Genentech Inc | Dosing for treatment with anti-cd20/anti-cd3 bispecific antibodies and anti-cd79b antibody drug conjugates. |
JP2023549316A (en) | 2020-11-16 | 2023-11-24 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Combination therapy with CD40 agonists targeting FAP |
EP4255928A1 (en) | 2020-12-02 | 2023-10-11 | GlaxoSmithKline Intellectual Property Development Limited | Il-7 binding proteins and their use in medical therapy |
TW202237638A (en) | 2020-12-09 | 2022-10-01 | 日商武田藥品工業股份有限公司 | Compositions of guanylyl cyclase c (gcc) antigen binding agents and methods of use thereof |
TW202237639A (en) | 2020-12-09 | 2022-10-01 | 日商武田藥品工業股份有限公司 | Compositions of guanylyl cyclase c (gcc) antigen binding agents and methods of use thereof |
WO2022132904A1 (en) | 2020-12-17 | 2022-06-23 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Human monoclonal antibodies targeting sars-cov-2 |
WO2022148732A1 (en) | 2021-01-06 | 2022-07-14 | F. Hoffmann-La Roche Ag | Combination therapy employing a pd1-lag3 bispecific antibody and a cd20 t cell bispecific antibody |
WO2022152656A1 (en) | 2021-01-12 | 2022-07-21 | F. Hoffmann-La Roche Ag | Split antibodies which bind to cancer cells and target radionuclides to said cells |
KR20230131205A (en) | 2021-01-13 | 2023-09-12 | 에프. 호프만-라 로슈 아게 | combination therapy |
WO2022162587A1 (en) | 2021-01-27 | 2022-08-04 | Centre Hospitalier Universitaire Vaudois (C.H.U.V.) | Anti-sars-cov-2 antibodies and use thereof in the treatment of sars-cov-2 infection |
WO2022162203A1 (en) | 2021-01-28 | 2022-08-04 | Vaccinvent Gmbh | Method and means for modulating b-cell mediated immune responses |
KR20230147099A (en) | 2021-01-28 | 2023-10-20 | 백신벤트 게엠베하 | METHOD AND MEANS FOR MODULATING B-CELL MEDIATED IMMUNE RESPONSES |
CN117120084A (en) | 2021-01-28 | 2023-11-24 | 维肯芬特有限责任公司 | Methods and means for modulating B cell mediated immune responses |
JP2024509695A (en) | 2021-02-03 | 2024-03-05 | ジェネンテック, インコーポレイテッド | Multispecific binding proteolysis platform and methods of use |
EP4291306A1 (en) | 2021-02-09 | 2023-12-20 | University of Georgia Research Foundation, Inc. | Human monoclonal antibodies against pneumococcal antigens |
CA3209136A1 (en) | 2021-02-09 | 2022-08-18 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Antibodies targeting the spike protein of coronaviruses |
WO2022172085A2 (en) | 2021-02-15 | 2022-08-18 | Takeda Pharmaceutical Company Limited | Cell therapy compositions and methods for modulating tgf-b signaling |
GB202102396D0 (en) | 2021-02-19 | 2021-04-07 | Adc Therapeutics Sa | Molecular adjuvant |
WO2022180145A2 (en) | 2021-02-26 | 2022-09-01 | Bayer Aktiengesellschaft | Inhibitors of il-11 or il-11ra for use in the treatment of abnormal uterine bleeding |
EP4301467A1 (en) | 2021-03-01 | 2024-01-10 | Xilio Development, Inc. | Combination of ctla4 and pd1/pdl1 antibodies for treating cancer |
EP4301781A1 (en) | 2021-03-01 | 2024-01-10 | Xilio Development, Inc. | Combination of masked ctla4 and pd1/pdl1 antibodies for treating cancer |
JP2024509169A (en) | 2021-03-03 | 2024-02-29 | ソレント・セラピューティクス・インコーポレイテッド | Antibody-drug conjugates including anti-BCMA antibodies |
JP2024512324A (en) | 2021-03-05 | 2024-03-19 | ジーオー セラピューティクス,インコーポレイテッド | Anti-glycoCD44 antibodies and their uses |
TW202302646A (en) | 2021-03-05 | 2023-01-16 | 美商當康生物科技有限公司 | Anti-vista constructs 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 |
AU2022232007A1 (en) | 2021-03-12 | 2023-10-26 | Janssen Biotech, Inc. | Method of treating psoriatic arthritis patients with inadequate response to tnf therapy with anti-il23 specific antibody |
CA3212729A1 (en) | 2021-03-12 | 2022-09-15 | Janssen Biotech, Inc. | Safe and effective method of treating psoriatic arthritis with anti-il23 specific antibody |
JP2024511970A (en) | 2021-03-15 | 2024-03-18 | ジェネンテック, インコーポレイテッド | Compositions and methods for the treatment of lupus nephritis |
WO2022197877A1 (en) | 2021-03-19 | 2022-09-22 | Genentech, Inc. | Methods and compositions for time delayed bio-orthogonal release of cytotoxic agents |
WO2022204724A1 (en) | 2021-03-25 | 2022-09-29 | Dynamicure Biotechnology Llc | Anti-igfbp7 constructs and uses thereof |
JP2024512633A (en) | 2021-03-30 | 2024-03-19 | バイエル・アクチエンゲゼルシヤフト | Anti-SEMA3A antibody and its uses |
AR125344A1 (en) | 2021-04-15 | 2023-07-05 | Chugai Pharmaceutical Co Ltd | ANTI-C1S ANTIBODY |
IL307501A (en) | 2021-04-19 | 2023-12-01 | Hoffmann La Roche | Modified mammalian cells |
JP2024514281A (en) | 2021-04-23 | 2024-04-01 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Prevention or reduction of adverse effects related to NK cell-engaging agents |
TW202243689A (en) | 2021-04-30 | 2022-11-16 | 瑞士商赫孚孟拉羅股份公司 | Dosing for combination treatment with anti-cd20/anti-cd3 bispecific antibody and anti-cd79b antibody drug conjugate |
TW202310876A (en) | 2021-05-12 | 2023-03-16 | 美商建南德克公司 | Methods of using anti-cd79b immunoconjugates to treat diffuse large b-cell lymphoma |
TW202306993A (en) | 2021-05-14 | 2023-02-16 | 美商建南德克公司 | Agonists of trem2 |
WO2022243261A1 (en) | 2021-05-19 | 2022-11-24 | F. Hoffmann-La Roche Ag | Agonistic cd40 antigen binding molecules targeting cea |
WO2022246259A1 (en) | 2021-05-21 | 2022-11-24 | Genentech, Inc. | Modified cells for the production of a recombinant product of interest |
CN117396513A (en) | 2021-05-28 | 2024-01-12 | 葛兰素史密斯克莱知识产权发展有限公司 | Combination therapy for the treatment of cancer |
AR126009A1 (en) | 2021-06-02 | 2023-08-30 | Hoffmann La Roche | CD28 ANTIGEN-BINDING AGONIST MOLECULES THAT TARGET EPCAM |
EP4155321A1 (en) | 2021-06-04 | 2023-03-29 | Chugai Seiyaku Kabushiki Kaisha | Anti-ddr2 antibodies and uses thereof |
KR20240019109A (en) | 2021-06-09 | 2024-02-14 | 에프. 호프만-라 로슈 아게 | Combination of a specific BRAF inhibitor (Paradox Break) and a PD-1 axis binding antagonist for use in the treatment of cancer |
IL308597A (en) | 2021-06-11 | 2024-01-01 | Genentech Inc | Method for treating chronic obstructive pulmonary disease with an st2 antagonist |
WO2022263638A1 (en) | 2021-06-17 | 2022-12-22 | Centre Hospitalier Universitaire Vaudois (C.H.U.V.) | Anti-sars-cov-2 antibodies and use thereof in the treatment of sars-cov-2 infection |
WO2022266660A1 (en) | 2021-06-17 | 2022-12-22 | Amberstone Biosciences, Inc. | Anti-cd3 constructs and uses thereof |
IL308633A (en) | 2021-06-25 | 2024-01-01 | Chugai Pharmaceutical Co Ltd | Use of anti-ctla-4 antibody |
CN117616123A (en) | 2021-06-25 | 2024-02-27 | 中外制药株式会社 | anti-CTLA-4 antibodies |
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 |
US20230038355A1 (en) | 2021-07-09 | 2023-02-09 | Janssen Biotech, Inc. | Manufacturing Methods for Producing Anti-IL12/IL23 Antibody Compositions |
TW202306985A (en) | 2021-07-12 | 2023-02-16 | 美商建南德克公司 | Structures for reducing antibody-lipase binding |
CN115812082A (en) | 2021-07-14 | 2023-03-17 | 舒泰神(北京)生物制药股份有限公司 | Antibody specifically recognizing CD40 and application thereof |
US20230049152A1 (en) | 2021-07-14 | 2023-02-16 | Genentech, Inc. | Anti-c-c motif chemokine receptor 8 (ccr8) antibodies and methods of use |
WO2023004386A1 (en) | 2021-07-22 | 2023-01-26 | Genentech, Inc. | Brain targeting compositions and methods of use thereof |
CA3219606A1 (en) | 2021-07-22 | 2023-01-26 | F. Hoffmann-La Roche Ag | Heterodimeric fc domain antibodies |
WO2023012147A1 (en) | 2021-08-03 | 2023-02-09 | F. Hoffmann-La Roche Ag | Bispecific antibodies and methods of use |
AU2022324456A1 (en) | 2021-08-05 | 2024-02-15 | Go Therapeutics, Inc. | Anti-glyco-muc4 antibodies and their uses |
US20230099756A1 (en) | 2021-08-07 | 2023-03-30 | Genentech, Inc. | Methods of using anti-cd79b immunoconjugates to treat diffuse large b-cell lymphoma |
IL310698A (en) | 2021-08-13 | 2024-04-01 | Glaxosmithkline Intellectual Property Development Ltd | Cytotoxicity targeting chimeras for ccr2-expressing cells |
WO2023019239A1 (en) | 2021-08-13 | 2023-02-16 | Genentech, Inc. | Dosing for anti-tryptase antibodies |
CA3227835A1 (en) | 2021-08-13 | 2023-02-16 | Peiling CHEN | Cytotoxicity targeting chimeras |
WO2023021055A1 (en) | 2021-08-19 | 2023-02-23 | F. Hoffmann-La Roche Ag | Multivalent anti-variant fc-region antibodies and methods of use |
CA3229448A1 (en) | 2021-08-23 | 2023-03-02 | Immunitas Therapeutics, Inc. | Anti-cd161 antibodies and uses thereof |
TW202328177A (en) | 2021-08-27 | 2023-07-16 | 美商建南德克公司 | 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 |
WO2023034571A1 (en) | 2021-09-03 | 2023-03-09 | Go Therapeutics, Inc. | Anti-glyco-lamp1 antibodies and their uses |
CA3230934A1 (en) | 2021-09-03 | 2023-03-09 | Go Therapeutics, Inc. | Anti-glyco-cmet antibodies and their uses |
AU2022345251A1 (en) | 2021-09-17 | 2024-03-28 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Synthetic humanized llama nanobody library and use thereof to identify sars-cov-2 neutralizing antibodies |
TW202321308A (en) | 2021-09-30 | 2023-06-01 | 美商建南德克公司 | Methods for treatment of hematologic cancers using anti-tigit antibodies, anti-cd38 antibodies, and pd-1 axis binding antagonists |
WO2023057893A1 (en) | 2021-10-05 | 2023-04-13 | Glaxosmithkline Intellectual Property Development Limited | Combination therapies for treating cancer |
WO2023073615A1 (en) | 2021-10-29 | 2023-05-04 | Janssen Biotech, Inc. | Methods of treating crohn's disease with anti-il23 specific antibody |
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 |
US20230151087A1 (en) | 2021-11-15 | 2023-05-18 | Janssen Biotech, Inc. | Methods of Treating Crohn's Disease with Anti-IL23 Specific Antibody |
WO2023088959A1 (en) | 2021-11-16 | 2023-05-25 | Ac Immune Sa | Novel molecules for therapy and diagnosis |
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 |
US20230159633A1 (en) | 2021-11-23 | 2023-05-25 | Janssen Biotech, Inc. | Method of Treating Ulcerative Colitis with Anti-IL23 Specific Antibody |
WO2023094569A1 (en) | 2021-11-26 | 2023-06-01 | F. Hoffmann-La Roche Ag | Combination therapy of anti-tyrp1/anti-cd3 bispecific antibodies and tyrp1-specific antibodies |
AR127887A1 (en) | 2021-12-10 | 2024-03-06 | F Hoffmann La Roche Ag | ANTIBODIES THAT BIND CD3 AND PLAP |
AR128031A1 (en) | 2021-12-20 | 2024-03-20 | F Hoffmann La Roche Ag | ANTI-LTBR AGONIST ANTIBODIES AND BISPECIFIC ANTIBODIES THAT COMPRISE THEM |
WO2023131901A1 (en) | 2022-01-07 | 2023-07-13 | Johnson & Johnson Enterprise Innovation Inc. | Materials and methods of il-1beta binding proteins |
WO2023141445A1 (en) | 2022-01-19 | 2023-07-27 | Genentech, Inc. | Anti-notch2 antibodies and conjugates and methods of use |
WO2023154824A1 (en) | 2022-02-10 | 2023-08-17 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Human monoclonal antibodies that broadly target coronaviruses |
TW202342519A (en) | 2022-02-16 | 2023-11-01 | 瑞士商Ac 免疫有限公司 | Humanized anti-tdp-43 binding molecules and uses thereof |
TW202342520A (en) | 2022-02-18 | 2023-11-01 | 美商樂天醫藥生技股份有限公司 | Anti-programmed death-ligand 1 (pd-l1) antibody molecules, encoding polynucleotides, and methods of use |
WO2023161875A1 (en) | 2022-02-25 | 2023-08-31 | Glaxosmithkline Intellectual Property Development Limited | Cytotoxicity targeting chimeras for prostate specific membrane antigen-expressing cells |
WO2023161874A1 (en) | 2022-02-25 | 2023-08-31 | Glaxosmithkline Intellectual Property Development Limited | Cytotoxicity targeting chimeras for c-c chemokine receptor 2-expressing cells |
WO2023161878A1 (en) | 2022-02-25 | 2023-08-31 | Glaxosmithkline Intellectual Property Development Limited | Cytotoxicity targeting chimeras for folate receptor-expressing cells |
WO2023161881A1 (en) | 2022-02-25 | 2023-08-31 | Glaxosmithkline Intellectual Property Development Limited | Cytotoxicity targeting chimeras for ccr2-expressing cells |
WO2023161876A1 (en) | 2022-02-25 | 2023-08-31 | Glaxosmithkline Intellectual Property Development Limited | Cytotoxicity targeting chimeras for cxcr3-expressing cells |
WO2023161877A1 (en) | 2022-02-25 | 2023-08-31 | Glaxosmithkline Intellectual Property Development Limited | Cytotoxicity targeting chimeras for integrin avb6-expressing cells |
WO2023161879A1 (en) | 2022-02-25 | 2023-08-31 | Glaxosmithkline Intellectual Property Development Limited | Cytotoxicity targeting chimeras for fibroblast activation protein-expressing cells |
WO2023173026A1 (en) | 2022-03-10 | 2023-09-14 | Sorrento Therapeutics, Inc. | Antibody-drug conjugates and uses thereof |
US20230414750A1 (en) | 2022-03-23 | 2023-12-28 | Hoffmann-La Roche Inc. | Combination treatment of an anti-cd20/anti-cd3 bispecific antibody and chemotherapy |
WO2023192827A1 (en) | 2022-03-26 | 2023-10-05 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Bispecific antibodies to hiv-1 env and their use |
WO2023186756A1 (en) | 2022-03-28 | 2023-10-05 | F. Hoffmann-La Roche Ag | Interferon gamma variants and antigen binding molecules comprising these |
WO2023192881A1 (en) | 2022-03-28 | 2023-10-05 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Neutralizing antibodies to hiv-1 env and their use |
WO2023187707A1 (en) | 2022-03-30 | 2023-10-05 | Janssen Biotech, Inc. | Method of treating mild to moderate psoriasis with il-23 specific antibody |
GB202204813D0 (en) | 2022-04-01 | 2022-05-18 | Bradcode Ltd | Human monoclonal antibodies and methods of use thereof |
WO2023191816A1 (en) | 2022-04-01 | 2023-10-05 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2023194565A1 (en) | 2022-04-08 | 2023-10-12 | Ac Immune Sa | Anti-tdp-43 binding molecules |
WO2023198727A1 (en) | 2022-04-13 | 2023-10-19 | F. Hoffmann-La Roche Ag | Pharmaceutical compositions of anti-cd20/anti-cd3 bispecific antibodies and methods of use |
WO2023201299A1 (en) | 2022-04-13 | 2023-10-19 | Genentech, Inc. | Pharmaceutical compositions of therapeutic proteins and methods of use |
WO2023203177A1 (en) | 2022-04-20 | 2023-10-26 | Kantonsspital St. Gallen | Antibodies or antigen-binding fragments pan-specifically binding to gremlin-1 and gremlin-2 and uses thereof |
WO2023209177A1 (en) | 2022-04-29 | 2023-11-02 | Astrazeneca Uk Limited | Sars-cov-2 antibodies and methods of using the same |
WO2023212304A1 (en) | 2022-04-29 | 2023-11-02 | 23Andme, Inc. | Antigen binding proteins |
WO2023215737A1 (en) | 2022-05-03 | 2023-11-09 | Genentech, Inc. | Anti-ly6e antibodies, immunoconjugates, and uses thereof |
TW202402810A (en) | 2022-05-11 | 2024-01-16 | 瑞士商赫孚孟拉羅股份公司 | Antibody that binds to vegf-a and il6 and methods of use |
WO2023219613A1 (en) | 2022-05-11 | 2023-11-16 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2023223265A1 (en) | 2022-05-18 | 2023-11-23 | Janssen Biotech, Inc. | Method for evaluating and treating psoriatic arthritis with il23 antibody |
WO2023227641A1 (en) | 2022-05-27 | 2023-11-30 | Glaxosmithkline Intellectual Property Development Limited | Use of tnf-alpha binding proteins and il-7 binding proteins in medical treatment |
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 |
WO2023237706A2 (en) | 2022-06-08 | 2023-12-14 | Institute For Research In Biomedicine (Irb) | Cross-specific antibodies, uses and methods for discovery thereof |
WO2024015897A1 (en) | 2022-07-13 | 2024-01-18 | 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 |
WO2024020432A1 (en) | 2022-07-19 | 2024-01-25 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
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 |
WO2024030829A1 (en) | 2022-08-01 | 2024-02-08 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Monoclonal antibodies that bind to the underside of influenza viral neuraminidase |
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 |
WO2024042112A1 (en) | 2022-08-25 | 2024-02-29 | Glaxosmithkline Intellectual Property Development Limited | Antigen binding proteins and uses thereof |
WO2024044779A2 (en) | 2022-08-26 | 2024-02-29 | Juno Therapeutics, Inc. | Antibodies and chimeric antigen receptors specific for delta-like ligand 3 (dll3) |
WO2024049949A1 (en) | 2022-09-01 | 2024-03-07 | Genentech, Inc. | Therapeutic and diagnostic methods for bladder cancer |
WO2024054822A1 (en) | 2022-09-07 | 2024-03-14 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Engineered sars-cov-2 antibodies with increased neutralization breadth |
WO2024054929A1 (en) | 2022-09-07 | 2024-03-14 | Dynamicure Biotechnology Llc | Anti-vista constructs and uses thereof |
WO2024064826A1 (en) | 2022-09-22 | 2024-03-28 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Neutralizing antibodies to plasmodium falciparum circumsporozoite protein and their use |
WO2024068996A1 (en) | 2022-09-30 | 2024-04-04 | Centre Hospitalier Universitaire Vaudois (C.H.U.V.) | Anti-sars-cov-2 antibodies and use thereof in the treatment of sars-cov-2 infection |
Citations (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4215051A (en) * | 1979-08-29 | 1980-07-29 | Standard Oil Company (Indiana) | Formation, purification and recovery of phthalic anhydride |
US4946778A (en) * | 1987-09-21 | 1990-08-07 | Genex Corporation | Single polypeptide chain binding molecules |
US4978745A (en) * | 1987-11-23 | 1990-12-18 | Centocor, Inc. | Immunoreactive heterochain antibodies |
US5047335A (en) * | 1988-12-21 | 1991-09-10 | The Regents Of The University Of Calif. | Process for controlling intracellular glycosylation of proteins |
US5529922A (en) * | 1989-05-25 | 1996-06-25 | Sloan-Kettering Institute For Cancer Research | Anti-idiotypic monoclonal antibody that induces an immune response against the ganglioside GD3 and hybridoma producing said antibody |
US5547933A (en) * | 1983-12-13 | 1996-08-20 | Kirin-Amgen, Inc. | Production of erythropoietin |
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 |
US5753229A (en) * | 1991-09-25 | 1998-05-19 | Mordoh; Jose | Monoclonal antibodies reactive with tumor proliferating cells |
US5776456A (en) * | 1992-11-13 | 1998-07-07 | Idec Pharmaceuticals Corporation | Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma |
US5939068A (en) * | 1995-06-07 | 1999-08-17 | Idec Pharmaceuticals Corporation | Neutralizing high affinity human monoclonal antibodies specific to RSV F-protein and methods for their manufacture and therapeutic use thereof |
US5952203A (en) * | 1997-04-11 | 1999-09-14 | The University Of British Columbia | Oligosaccharide synthesis using activated glycoside derivative, glycosyl transferase and catalytic amount of nucleotide phosphate |
US5958403A (en) * | 1992-02-28 | 1999-09-28 | Beth Israel Hospital Association | Methods and compounds for prevention of graft rejection |
US6153433A (en) * | 1995-11-17 | 2000-11-28 | Takara Shuzo Co., Ltd. | Inhibitor for viral replication |
US6183744B1 (en) * | 1997-03-24 | 2001-02-06 | Immunomedics, Inc. | Immunotherapy of B-cell malignancies using anti-CD22 antibodies |
US6602684B1 (en) * | 1998-04-20 | 2003-08-05 | Glycart Biotechnology Ag | Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity |
US20030175884A1 (en) * | 2001-08-03 | 2003-09-18 | Pablo Umana | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
Family Cites Families (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE4028800A1 (en) | 1990-09-11 | 1992-03-12 | Behringwerke Ag | GENETIC SIALYLATION OF GLYCOPROTEINS |
US5558865A (en) | 1991-08-22 | 1996-09-24 | Nissin Shokuhin Kabushiki Kaisha | HIV immunotherapeutics |
SG44845A1 (en) | 1993-01-12 | 1997-12-19 | Biogen Inc | Recombitant anti-vla4 antibody molecules |
JPH09509847A (en) | 1994-03-09 | 1997-10-07 | アボツト・ラボラトリーズ | Human emulsified milk |
GB9603256D0 (en) * | 1996-02-16 | 1996-04-17 | Wellcome Found | Antibodies |
WO1998006855A1 (en) | 1996-08-16 | 1998-02-19 | The Texas A & M University System | Compositions and methods for delivery of nucleic acids to hepatocytes |
US6306393B1 (en) | 1997-03-24 | 2001-10-23 | Immunomedics, Inc. | Immunotherapy of B-cell malignancies using anti-CD22 antibodies |
US6946292B2 (en) | 2000-10-06 | 2005-09-20 | Kyowa Hakko Kogyo Co., Ltd. | Cells producing antibody compositions with increased antibody dependent cytotoxic activity |
JP2005500018A (en) * | 2001-04-02 | 2005-01-06 | アイデック ファーマスーティカルズ コーポレイション | Recombinant antibody coexpressed with GnTIII |
PL224786B1 (en) | 2003-01-22 | 2017-01-31 | Glycart Biotechnology Ag | Fusion constructs and use of same to produce antibodies with increased fc receptor binding affinity and effector function |
ME01775B (en) | 2003-11-05 | 2011-02-28 | Glycart Biotechnology Ag | Cd20 antibodies with increased fc receptor binding affinity and effector function |
PL1871805T3 (en) | 2005-02-07 | 2020-03-31 | Roche Glycart Ag | Antigen binding molecules that bind egfr, vectors encoding same, and uses thereof |
NO345919B1 (en) | 2005-08-26 | 2021-10-18 | Roche Glycart Ag | Modified antigen-binding molecules with altered cell signaling activity |
AR062223A1 (en) | 2006-08-09 | 2008-10-22 | Glycart Biotechnology Ag | MOLECULES OF ADHESION TO THE ANTIGEN THAT ADHER TO EGFR, VECTORS THAT CODE THEM, AND THEIR USES OF THESE |
SG178567A1 (en) | 2009-08-31 | 2012-04-27 | Roche Glycart Ag | Affinity-matured humanized anti cea monoclonal antibodies |
-
2002
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- 2002-08-05 WO PCT/US2002/024739 patent/WO2003011878A2/en active Application Filing
-
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- 2004-02-02 IL IL160170A patent/IL160170A/en unknown
- 2004-02-02 NO NO20040453A patent/NO332457B1/en not_active IP Right Cessation
-
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- 2005-08-09 US US11/199,232 patent/US8021856B2/en not_active Expired - Fee Related
-
2008
- 2008-12-25 JP JP2008331038A patent/JP2009114201A/en active Pending
-
2011
- 2011-08-02 US US13/196,724 patent/US8999324B2/en not_active Expired - Fee Related
-
2015
- 2015-03-23 US US14/665,191 patent/US9321843B2/en not_active Expired - Fee Related
-
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- 2016-03-24 US US15/080,020 patent/US9631023B2/en not_active Expired - Fee Related
Patent Citations (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4215051A (en) * | 1979-08-29 | 1980-07-29 | Standard Oil Company (Indiana) | Formation, purification and recovery of phthalic anhydride |
US5547933A (en) * | 1983-12-13 | 1996-08-20 | Kirin-Amgen, Inc. | Production of erythropoietin |
US4946778A (en) * | 1987-09-21 | 1990-08-07 | Genex Corporation | Single polypeptide chain binding molecules |
US4978745A (en) * | 1987-11-23 | 1990-12-18 | Centocor, Inc. | Immunoreactive heterochain antibodies |
US5047335A (en) * | 1988-12-21 | 1991-09-10 | The Regents Of The University Of Calif. | Process for controlling intracellular glycosylation of proteins |
US5529922A (en) * | 1989-05-25 | 1996-06-25 | Sloan-Kettering Institute For Cancer Research | Anti-idiotypic monoclonal antibody that induces an immune response against the ganglioside GD3 and hybridoma producing said antibody |
US5753229A (en) * | 1991-09-25 | 1998-05-19 | Mordoh; Jose | Monoclonal antibodies reactive with tumor proliferating cells |
US5958403A (en) * | 1992-02-28 | 1999-09-28 | Beth Israel Hospital Association | Methods and compounds for prevention of graft rejection |
US5776456A (en) * | 1992-11-13 | 1998-07-07 | Idec Pharmaceuticals Corporation | Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma |
US5843439A (en) * | 1992-11-13 | 1998-12-01 | Anderson; Darrell R. | Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma |
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 |
US5939068A (en) * | 1995-06-07 | 1999-08-17 | Idec Pharmaceuticals Corporation | Neutralizing high affinity human monoclonal antibodies specific to RSV F-protein and methods for their manufacture and therapeutic use thereof |
US6153433A (en) * | 1995-11-17 | 2000-11-28 | Takara Shuzo Co., Ltd. | Inhibitor for viral replication |
US6183744B1 (en) * | 1997-03-24 | 2001-02-06 | Immunomedics, Inc. | Immunotherapy of B-cell malignancies using anti-CD22 antibodies |
US5952203A (en) * | 1997-04-11 | 1999-09-14 | The University Of British Columbia | Oligosaccharide synthesis using activated glycoside derivative, glycosyl transferase and catalytic amount of nucleotide phosphate |
US6602684B1 (en) * | 1998-04-20 | 2003-08-05 | Glycart Biotechnology Ag | Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity |
US20040072290A1 (en) * | 1998-04-20 | 2004-04-15 | Glycart Biotechnology Ag | Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity |
US20050074843A1 (en) * | 1998-04-20 | 2005-04-07 | Pablo Umana | Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity |
US20030175884A1 (en) * | 2001-08-03 | 2003-09-18 | Pablo Umana | Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity |
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US20090060911A1 (en) * | 2000-04-13 | 2009-03-05 | The Rockefeller University | Enhancement of antibody-mediated immune responses |
US20110052584A1 (en) * | 2000-04-13 | 2011-03-03 | The Rockefeller University | Method of enhancement of cytotoxicity in antibody mediated immune responses |
US20050123546A1 (en) * | 2003-11-05 | 2005-06-09 | Glycart Biotechnology Ag | Antigen binding molecules with increased Fc receptor binding affinity and effector function |
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US9296820B2 (en) | 2003-11-05 | 2016-03-29 | Roche Glycart Ag | Polynucleotides encoding anti-CD20 antigen binding molecules with increased Fc receptor binding affinity and effector function |
US20100081195A1 (en) * | 2008-09-26 | 2010-04-01 | Eureka Therapeutics, Inc. | Modified Host Cells and Uses Thereof |
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US8025879B2 (en) | 2008-09-26 | 2011-09-27 | Eureka Therapeutics, Inc. | Modified glycoproteins and uses thereof |
US8080415B2 (en) | 2008-09-26 | 2011-12-20 | Eureka Therapeutics, Inc. | Modified host cells and uses thereof |
US8084222B2 (en) | 2008-09-26 | 2011-12-27 | Eureka Therapeutics, Inc. | Methods for generating host cells |
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