US20080299618A1 - Single domain ligands, receptors comprising said ligands, methods for their production and use of said ligands and receptors - Google Patents
Single domain ligands, receptors comprising said ligands, methods for their production and use of said ligands and receptors Download PDFInfo
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- US20080299618A1 US20080299618A1 US12/127,237 US12723708A US2008299618A1 US 20080299618 A1 US20080299618 A1 US 20080299618A1 US 12723708 A US12723708 A US 12723708A US 2008299618 A1 US2008299618 A1 US 2008299618A1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/655—Somatostatins
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
- C07K16/461—Igs containing Ig-regions, -domains or -residues form different species
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
Definitions
- the present invention relates to single domain ligands derived from molecules in the immunoglobulin (Ig) superfamily, receptors comprising at least one such ligand, methods for cloning, amplifying and expressing DNA sequences encoding such ligands, methods for the use of said DNA sequences in the production of Ig-type molecules and said ligands or receptors, and the use of said ligands or receptors in therapy, diagnosis or catalysis.
- Ig immunoglobulin
- FIG. 1 shows a schematic representation of the unrearranged and rearranged heavy and light chain variable genes and the location of the primers.
- FIG. 2 shows a schematic representation of the M13-VHPCR1 vector and a cloning scheme for amplified heavy chain variable domains.
- FIG. 3 shows the sequence of the Ig variable region derived sequences in M13-VHPCR1.
- FIG. 4 shows a schematic representation of the M13-VKPCR1 vector and a cloning scheme for light chain variable domains.
- FIG. 5 shows the sequence of the Ig variable region derived sequences in M13-VKPCR1.
- FIG. 6 shows the nucleotide sequences of the heavy and light chain variable domain encoding sequences of MAb MBr1.
- FIG. 7 shows a schematic representation of the pSV-gpt vector (also known as ⁇ -Lys 30) which contains a variable region cloned as a HindIII-BamHI fragment, which is excised on introducing the new variable region.
- the gene for human IgG1 has also been engineered to remove a BamHI site, such that the BamHI site in the vector is unique.
- FIG. 8 shows a schematic representation of the pSV-hygro vector (also known as ⁇ -Lys 17). It is derived from pSV gpt vector with the gene encoding mycophenolic acid replaced by a gene coding for hygromycin resistance. The construct contains a variable gene cloned as a HindIII-BamHI fragment which is excised on introducing the new variable region. The gene for human C ⁇ has also been engineered to remove a BamHI site, such that the BamHI site in the vector is unique.
- FIG. 9 shows the assembly of the mouse: human MBr1 chimeric antibody.
- FIGS. 10 a - 10 b shows encoded amino acid sequences of 48 mouse rearranged VH genes.
- FIG. 11 shows encoded amino acid sequences of human rearranged VH genes.
- FIG. 12 shows encoded amino acid sequences of unrearranged human VH genes.
- FIG. 13 shows the sequence of part of the plasmid pSW1: essentially the sequence of a pectate lyase leader linked to VHLYS in pSW1 and cloned as an SphI-EcoRI fragment into pUC19 and the translation of the open reading frame encoding the pectate lyase leader-VHLYS polypeptide being shown.
- FIGS. 14 a - 14 b shows the sequence of part of the plasmid pSW2: essentially the sequence of a pectate lyase leader linked to VHLYS and to VKLYS, and cloned as an SphI-EcoRI-EcoRI fragment into pUC19 and the translation of open reading frames encoding the pectate lyase leader-VHLYS and pectate lyase leader-VKLYS polypeptides being shown.
- FIG. 15 shows the sequence of part of the plasmid pSW1HPOLYMYC which is based on pSW1 and in which a polylinker sequence has replaced the variable domain of VHLYS, and acts as a cloning site for amplified VH genes, and a peptide tag is introduced at the C-terminal end.
- FIG. 16 shows the encoded amino acid sequences of two VH domains derived from mouse spleen and having lysozyme binding activity, and compared with the VH domain of the D1,3 antibody.
- the arrows mark the points of difference between the two VH domains.
- FIG. 17 shows the encoded amino acid sequence of a VH domain derived from human peripheral blood lymphocytes and having lysozyme binding activity.
- FIG. 18 shows a scheme for generating and cloning mutants of the VHLYS gene, which is compared with the scheme for cloning natural repertoires of VH genes.
- FIG. 19 shows the sequence of part of the vector pSW2HPOLY.
- FIG. 20 shows the sequence of part of the vector pSW3 which encodes the two linked VHLYS domains.
- FIGS. 21 a - 21 c shows the sequence of the VHLYS domain and pelB leader sequence fused to the alkaline phosphatase gene.
- FIG. 22 shows the sequence of the vector pSW1VHLYS-VKPOLYMYC for expression of a repertoire of V ⁇ light chain variable domains in association with the VHLYS domain.
- FIG. 23 shows the sequence of VH domain which is secreted at high levels from E. coli . The differences with VHLYS domain are marked.
- the present invention relates to single domain ligands derived from molecules in the immunoglobulin (Ig) superfamily, receptors comprising at least one such ligand, methods for cloning, amplifying and expressing DNA sequences encoding such ligands, methods for the use of said DNA sequences in the production of Ig-type molecules and said ligands or receptors, and the use of said ligands or receptors in therapy, diagnosis or catalysis.
- Ig immunoglobulin
- the Ig superfamily includes not only the Igs themselves but also such molecules as receptors on lymphoid cells such as T lymphocytes.
- Immunoglobulins comprise at least one heavy and one light chain covalently bonded together. Each chain is divided into a number of domains. At the N-terminal end of each chain is a variable domain. The variable domains on the heavy and light chains fit together to form a binding site designed to receive a particular target molecule. In the case of Igs, the target molecules are antigens.
- T-cell receptors have two chains of equal size, the ⁇ and ⁇ chains, each consisting of two domains.
- variable domains on the ⁇ and ⁇ chains are believed to fit together to form a binding site for target molecules, in this case peptides presented by a histocompatibility antigen.
- the variable domains are so called because their amino acid sequences vary particularly from one molecule to another. This variation in sequence enables the molecules to recognize an extremely wide variety of target molecules.
- each variable domain comprises a number of areas of relatively conserved sequence and three areas of hypervariable sequence.
- the three hypervariable areas are generally known as complementarity determining regions (CDRs).
- Boss The Boss application also relates to the cloning and expression of chimeric antibodies.
- Chimeric antibodies are Ig-type molecules in which the variable domains from one Ig are fused to constant domains from another Ig.
- the variable domains are derived from an Ig from one species (often a mouse Ig) and the constant domains are derived from an Ig from a different species (often a human Ig).
- EP-A-0 125 023 (Genentech) relates to much the same subject as the Boss application, but also relates to the production by recombinant DNA technology of other variations of Ig-type molecules.
- EP-A-0 194 276 discloses not only chimeric antibodies of the type disclosed in the Boss application but also chimeric antibodies in which some or all of the constant domains have been replaced by non-Ig derived protein sequences.
- the heavy chain CH2 and CH3 domains may be replaced by protein sequences derived from an enzyme or a protein toxin.
- EP-A-0 239 400 discloses a different approach to the production of Ig molecules.
- this approach only the CDRs from a first type of Ig are grafted onto a second type of Ig in place of its normal CDRs.
- the Ig molecule thus produced is predominantly of the second type, since the CDRs form a relatively small part of the whole Ig.
- the CDRs are the parts which define the specificity of the Ig, the Ig molecule thus produced has its specificity derived from the first Ig.
- modified antibodies chimeric antibodies, CDR-grafted Igs, the altered antibodies described by Genentech, and fragments of such Igs such as F(ab′) 2 and Fv fragments are referred to herein as modified antibodies.
- MAbs monoclonal antibodies
- MAbs directed against cancer antigens have been produced. It is envisaged that these MAbs could be covalently attached or fused to toxins to provide “magic bullets” for use in cancer therapy. MAbs directed against normal tissue or cell surface antigens have also been produced. Labels can be attached to these so that they can be used for in vivo imaging.
- MAbs in therapy or in vivo diagnosis
- the vast majority of MAbs which are produced are of rodent, in particular mouse, origin. It is very difficult to produce human MAbs. Since most MAbs are derived from non-human species, they are antigenic in humans. Thus, administration of these MAbs to humans generally results in an anti-Ig response being mounted by the human. Such a response can interfere with therapy or diagnosis, for instance by destroying or clearing the antibody quickly, or can cause allergic reactions or immune complex hypersensitivity which has adverse effects on the patient.
- modified Igs have been proposed to ensure that the Ig administered to a patient is as “human” as possible, but still retains the appropriate specificity. It is therefore expected that modified Igs will be as effective as the MAb from which the specificity is derived but at the same time not very antigenic. Thus, it should be possible to use the modified Ig a reasonable number of times in a treatment or diagnosis regime.
- heavy chain variable domains are encoded by a “rearranged” gene which is built from three gene segments: an “unrearranged” VH gene (encoding the N-terminal three framework regions, first two complete CDRs and the first part of the third CDR), a diversity (DH)-segment (DH) (encoding the central portion of the third CDR) and a joining segment (JH) (encoding the last part of the third CDR and the fourth framework region).
- VH gene encoding the N-terminal three framework regions, first two complete CDRs and the first part of the third CDR
- DH diversity-segment
- JH joining segment
- light chain variable domains are encoded by an “unrearranged” VL gene and a JL gene.
- VL gene There are two types of light chains, kappa ( ⁇ ) or lambda ( ⁇ ), which are built respectively from unrearranged V ⁇ genes and J ⁇ segments, and from unrearranged V ⁇ genes and J ⁇ segments.
- Ig heavy chain variable domains can bind to antigen in a 1:1 ratio and with binding constants of equivalent magnitude to those of complete antibody molecules. In view of what was known up until now and in view of the assumptions made by those skilled in the art, this is highly surprising.
- a single domain ligand consisting of at least part of the variable domain of one chain of a molecule from the Ig superfamily.
- the ligand consists of the variable domain of an Ig light, or, most preferably, heavy chain.
- the ligand may be produced by any known technique, for instance by controlled cleavage of Ig superfamily molecules or by peptide synthesis. However, preferably the ligand is produced by recombinant DNA technology. For instance, the gene encoding the rearranged gene for a heavy chain variable domain may be produced, for instance by cloning or gene synthesis, and placed into a suitable expression vector. The expression vector is then used to transform a compatible host cell which is then cultured to allow the ligand to be expressed and, preferably, secreted.
- the gene for the ligand can be mutated to improve the properties of the expressed domain, for example to increase the yields of expression or the solubility of the ligand, to enable the ligand to bind better, or to introduce a second site for covalent attachment (by introducing chemically reactive residues such as cysteine and histidine) or non-covalent binding of other molecules.
- a second site for binding to serum components to prolong the residence time of the domains in the serum; or for binding to molecules with effector functions, such as components of complement, or receptors on the surfaces of cells.
- hydrophobic residues which would normally be at the interface of the heavy chain variable domain with the light chain variable domain could be mutated to more hydrophilic residues to improve solubility; residues in the CDR loops could be mutated to improve antigen binding; residues on the other loops or parts of the ⁇ -sheet could be mutated to introduce new binding activities. Mutations could include single point mutations, multiple point mutations or more extensive changes and could be introduced by any of a variety of recombinant DNA methods, for example gene synthesis, site directed mutagenesis or the polymerase chain reaction.
- the ligands of the present invention have equivalent binding affinity to that of complete Ig molecules
- the ligands can be used in many of the ways as are Ig molecules or fragments.
- Ig molecules have been used in therapy (such as in treating cancer, bacterial and viral diseases), in diagnosis (such as pregnancy testing), in vaccination (such as in producing anti-idiotypic antibodies which mimic antigens), in modulation of activities of hormones or growth factors, in detection, in biosensors and in catalysis.
- the small size of the ligands of the present invention may confer some advantages over complete antibodies, for example, in neutralizing the activity of low molecular weight drugs (such as dioxin) and allowing their filtration from the kidneys with drug attached; in penetrating tissues and tumors; in neutralizing viruses by binding to small conserved regions on the surfaces of viruses such as the “canyon” sites of viruses [16]; in high resolution epitope mapping of proteins; and in vaccination by ligands which mimic antigens.
- low molecular weight drugs such as dioxin
- the present invention also provides receptors comprising a ligand according to the first aspect of the invention linked to one or more of an effector molecule, a label, a surface, or one or more other ligands having the same or different specificity.
- a receptor comprising a ligand linked to an effector molecule may be of use in therapy.
- the effector molecule may be a toxin, such as ricin or pseudomonas exotoxin, an enzyme which is able to activate a prodrug, a binding partner or a radio-isotope.
- the radio-isotope may be directly linked to the ligand or may be attached thereto by a chelating structure which is directly linked to the ligand.
- Such ligands with attached isotopes are much smaller than those based on Fv fragments, and could penetrate tissues and access tumors more readily.
- a receptor comprising a ligand linked to a label may be of use in diagnosis.
- the label may be a heavy metal atom or a radio-isotope, in which case the receptor can be used for in vivo imaging using X-ray or other scanning apparatus.
- the metal atom or radio-isotope may be attached to the ligand either directly or via a chelating structure directly linked to the ligand.
- the label may be a heavy metal atom, a radio-isotope, an enzyme, a fluorescent or colored molecule or a protein or peptide tag which can be detected by an antibody, an antibody fragment or another protein.
- Such receptors would be used in any of the known diagnostic tests, such as ELISA or fluorescence-linked assays.
- a receptor comprising a ligand linked to a surface could be used for purification of other molecules by affinity chromatography.
- Linking of ligands to cells for example to the outer membrane proteins of E. coli or to hydrophobic tails which localize the ligands in the cell membranes, could allow a simple diagnostic test in which the bacteria or cells would agglutinate in the presence of molecules bearing multiple sites for binding the ligand(s).
- Receptors comprising at least two ligands can be used, for instance, in diagnostic tests.
- the first ligand will bind to a test antigen and the second ligand will bind to a reporter molecule, such as an enzyme, a fluorescent dye, a colored dye, a radio-isotope or a colored-, fluorescently- or radio-labelled protein.
- such receptors may be useful in increasing the binding to an antigen.
- the first ligand will bind to a first epitope of the antigen and the second ligand will bind to a second epitope.
- Such receptors may also be used for increasing the affinity and specificity of binding to different antigens in close proximity on the surface of cells.
- the first ligand will bind to the first antigen and the second epitope to the second antigen: strong binding will depend on the co-expression of the epitopes on the surface of the cell. This may be useful in therapy of tumors, which can have elevated expression of several surface markers. Further ligands could be added to further improve binding or specificity.
- the use of strings of ligands with the same or multiple specificities, creates a larger molecule which is less readily filtered from the circulation by the kidney.
- the use of strings of ligands may prove more effective than single ligands, due to repetition of the immunizing epitopes.
- such receptors with multiple ligands could include effector molecules or labels so that they can be used in therapy or diagnosis as described above.
- the ligand may be linked to the other part of the receptor by any suitable means, for instance by covalent or non-covalent chemical linkages.
- the receptor comprises a ligand and another protein molecule, it is preferred that they are produced by recombinant DNA technology as a fusion product. If necessary, a linker peptide sequence can be placed between the ligand and the other protein molecule to provide flexibility.
- the ligand is to be used for in vivo diagnosis or therapy in humans, it is humanized, for instance by CDR replacement as described in EP-A-0 239 400.
- a further problem with the production of ligands, and also receptors according to the invention and modified Igs, by recombinant DNA technology is the cloning of the variable domain encoding sequences from the hybridoma which produces the MAb from which the specificity is to be derived.
- This can be a relatively long method involving the production of a suitable probe, construction of a clone library from cDNA or genomic DNA, extensive probing of the clone library, and manipulation of any isolated clones to enable the cloning into a suitable expression vector. Due to the inherent variability of the DNA sequences encoding Ig variable domains, it has not previously been possible to avoid such time consuming work. It is therefore a further aim of the present invention to provide a method which enables substantially any sequence encoding an Ig superfamily molecule variable domain (ligand) to be cloned in a reasonable period of time.
- a method of cloning a sequence (the target sequence) which encodes at least part of the variable domain of an Ig superfamily molecule which method comprises:
- a forward and a back oligonucleotide primer annealing to the sample a forward and a back oligonucleotide primer, the forward primer being specific for a sequence at or adjacent the 3′ end of the sense strand of the target sequence, the back primer being specific for a sequence at or adjacent the 3′ end of the antisense strand of the target sequence, under conditions which allow the primers to hybridize to the nucleic acid at or adjacent the target sequence;
- the method of the present invention further includes the step (f) of repeating steps (c) to (e) on the denatured mixture a plurality of times.
- the method of the present invention is used to clone complete variable domains from Ig molecules, most preferably from Ig heavy chains.
- the method will produce a DNA sequence encoding a ligand according to the present invention.
- step (c) recited above the forward primer becomes annealed to the sense strand of the target sequence at or adjacent the 3′ end of the strand.
- the back primer becomes annealed to the antisense strand of the target sequence at or adjacent the 3′ end of the strand.
- the forward primer anneals at or adjacent the region of the ds nucleic acid which encodes the C-terminal end of the variable region or domain.
- the back primer anneals at or adjacent the region of the ds nucleic acid which encodes the N-terminal end of the variable domain.
- step (d) nucleotides are added onto the 3′ end of the forward and back primers in accordance with the sequence of the strand to which they are annealed. Primer extension will continue in this manner until stopped by the beginning of the denaturing step (e). It must therefore be ensured that step (d) is carried out for a long enough time to ensure that the primers are extended so that the extended strands totally overlap one another.
- step (e) the extended primers are separated from the ds nucleic acid.
- the ds nucleic acid can then serve again as a substrate to which further primers can anneal.
- the extended primers themselves have the necessary complementary sequences to enable the primers to anneal thereto.
- step (f) the amount of extended primers will increase exponentially so that at the end of the cycles there will be a large quantity of cDNA having sequences complementary to the sense and antisense strands of the target sequence.
- the method of the present invention will result in the accumulation of a large quantity of cDNA which can form ds cDNA encoding at least part of the variable domain.
- the forward and back primers may be provided as isolated oligonucleotides, in which case only two oligonucleotides will be used. However, alternatively the forward and back primers may each be supplied as a mixture of closely related oligonucleotides. For instance, it may be found that at a particular point in the sequence to which the primer is to anneal, there is the possibility of nucleotide variation. In this case a primer may be used for each possible nucleotide variation. Furthermore it may be possible to use two or more sets of “nested” primers in the method to enhance the specific cloning of variable region genes.
- RNA may be isolated in known manner from a cell or cell line which is known to produce Igs.
- mRNA may be separated from other RNA by oligo-dT chromatography.
- a complementary strand of cDNA may then be synthesized on the mRNA template, using reverse transcriptase and a suitable primer, to yield an RNA/DNA heteroduplex.
- a second strand of DNA can be made in one of several ways, for example, by priming with RNA fragments of the mRNA strand (made by incubating RNA/DNA heteroduplex with RNase H) and using DNA polymerase, or by priming with a synthetic oligodeoxynucleotide primer which anneals to the 3′ end of the first strand and using DNA polymerase. It has been found that the method of the present invention can be carried out using ds cDNA prepared in this way.
- a forward primer which anneals to a sequence in the CH1 domain (for a heavy chain variable domain) or the C ⁇ or C ⁇ domain (for a light chain variable domain). These will be located in close enough proximity to the target sequence to allow the sequence to be cloned.
- the back primer may be one which anneals to a sequence at the N-terminal end of the VH1, V ⁇ or V ⁇ domain.
- the back primer may consist of a plurality of primers having a variety of sequences designed to be complementary to the various families of VH1, V ⁇ or V ⁇ sequences known.
- the back primer may be a single primer having a consensus sequence derived from all the families of variable region genes.
- the method of the present invention can be carried out using genomic DNA. If genomic DNA is used, there is a very large amount of DNA present, including actual coding sequences, introns and untranslated sequences between genes. Thus, there is considerable scope for non-specific annealing under the conditions used. However, it has surprisingly been found that there is very little non-specific annealing. It is therefore unexpected that it has proved possible to clone the genes of Ig-variable domains from genomic DNA.
- genomic DNA may prove advantageous compared with use of mRNA, as the mRNA is readily degraded, and especially difficult to prepare from clinical samples of human tissue.
- the ds nucleic acid used in step (a) is genomic DNA.
- genomic DNA As the ds nucleic acid source, it will not be possible to use as the forward primer an oligonucleotide having a sequence complementary to a sequence in a constant domain. This is because, in genomic DNA, the constant domain genes are generally separated from the variable domain genes by a considerable number of base pairs. Thus, the site of annealing would be too remote from the sequence to be cloned.
- the method of the present invention can be used to clone both rearranged and unrearranged variable domain sequences from genomic DNA. It is known that in germ line genomic DNA the three genes, encoding the VH, DH and JH respectively, are separated from one another by considerable numbers of base pairs. On maturation of the immune response, these genes are rearranged so that the VH, DH and JH genes are fused together to provide the gene encoding the whole variable domain (see FIG. 1 ). By using a forward primer specific for a sequence at or adjacent the 3′ end of the sense strand of the genomic “unrearranged” VH gene, it is possible to clone the “unrearranged” VH gene alone, without also cloning the DH and JH genes. This can be of use in that it will then be possible to fuse the VH gene onto pre-cloned or synthetic DH and DH genes. In this way, rearrangement of the variable domain genes can be carried out in vitro.
- the oligonucleotide primers used in step (c) may be specifically designed for use with a particular target sequence. In this case, it will be necessary to sequence at least the 5′ and 3′ ends of the target sequence so that the appropriate oligonucleotides can be synthesized. However, the present inventors have discovered that it is not necessary to use such specifically designed primers. Instead, it is possible to use a species specific general primer or a mixture of such primers for annealing to each end of the target sequence. This is not particularly surprising as regards the 3′ end of the target sequence. It is known that this end of the variable domain encoding sequence leads into a segment encoding JH which is known to be relatively conserved. However, it was surprisingly discovered that, within a single species, the sequence at the 5′ end of the target sequence is sufficiently well conserved to enable a species specific general primer or a mixture thereof to be designed for the 5′ end of the target sequence.
- the two primers which are used are species specific general primers, whether used as single primers or as mixtures of primers. This greatly facilitates the cloning of any undetermined target sequence since it will avoid the need to carry out any sequencing on the target sequence in order to produce target sequence-specific primers.
- the method of this aspect of the invention provides a general method for cloning variable region or domain encoding sequences of a particular species.
- variable domain gene Once the variable domain gene has been cloned using the method described above, it may be directly inserted into an expression vector, for instance using the PCR reaction to paste the gene into a vector.
- each primer includes a sequence including a restriction enzyme recognition site.
- the sequence recognized by the restriction enzyme need not be in the part of the primer which anneals to the ds nucleic acid, but may be provided as an extension which does not anneal.
- the use of primers with restriction sites has the advantage that the DNA can be cut with at least one restriction enzyme which leaves 3′ or 5′ overhanging nucleotides. Such DNA is more readily cloned into the corresponding sites on the vectors than blunt end fragments taken directly from the method. The ds cDNA produced at the end of the cycles will thus be readily insertable into a cloning vector by use of the appropriate restriction enzymes.
- restriction sites is such that the ds cDNA is cloned directly into an expression vector, such that the ligand encoded by the gene is expressed.
- the restriction site is preferably located in the sequence which is annealed to the ds nucleic acid.
- the primers may not have a sequence exactly complementary to the target sequence to which it is to be annealed, for instance because of nucleotide variations or because of the introduction of a restriction enzyme recognition site, it may be necessary to adjust the conditions in the annealing mixture to enable the primers to anneal to the ds nucleic acid. This is well within the competence of the person skilled in the art and needs no further explanation.
- any DNA polymerase may be used.
- Such polymerases are known in the art and are available commercially. The conditions to be used with each polymerase are well known and require no further explanation here.
- the polymerase reaction will need to be carried out in the presence of the four nucleoside triphosphates. These and the polymerase enzyme may already be present in the sample or may be provided afresh for each cycle.
- the denaturing step (e) may be carried out, for instance, by heating the sample, by use of chaotropic agents, such as urea or guanidine, or by the use of changes in ionic strength or pH.
- chaotropic agents such as urea or guanidine
- denaturing is carried out by heating since this is readily reversible.
- thermostable DNA polymerase such as Taq polymerase, since this will not need replenishing at each cycle.
- a suitable cycle of heating comprises denaturation at about 95° C. for about 1 minute, annealing at from 30° C. to 65° C. for about 1 minute and primer extension at about 75° C. for about 2 minutes.
- the mixture after the final cycle is preferably held at about 60° C. for about 5 minutes.
- the product ds cDNA may be separated from the mixture for instance by gel electrophoresis using agarose gels.
- the ds cDNA may be used in unpurified form and inserted directly into a suitable cloning or expression vector by conventional methods. This will be particularly easy to accomplish if the primers include restriction enzyme recognition sequences.
- the method of the present invention may be used to make variations in the sequences encoding the variable domains.
- this may be achieved by using a mixture of related oligonucleotide primers as at least one of the primers.
- the primers are particularly variable in the middle of the primer and relatively conserved at the 5′ and 3′ ends.
- the ends of the primers are complementary to the framework regions of the variable domain, and the variable region in the middle of the primer covers all or part of a CDR.
- a forward primer is used in the area which forms the third CDR. If the method is carried out using such a mixture of oligonucleotides, the product will be a mixture of variable domain encoding sequences.
- variations in the sequence may be introduced by incorporating some mutagenic nucleotide triphosphates in step (d), such that point mutations are scattered throughout the target region.
- point mutations are introduced by performing a large number of cycles of amplification, as errors due to the natural error rate of the DNA polymerase are amplified, particularly when using high concentrations of nucleoside triphosphates.
- the method of this aspect of the present invention has the advantage that it greatly facilitates the cloning of variable domain encoding sequences directly from mRNA or genomic DNA. This in turn will facilitate the production of modified Ig-type molecules by any of the prior art methods referred to above. Further, target genes can be cloned from tissue samples containing antibody producing cells, and the genes can be sequenced. By doing this, it will be possible to look directly at the immune repertoire of a patient. This “fingerprinting” of a patient's immune repertoire could be of use in diagnosis, for instance of auto-immune diseases.
- a single set of primers is used in several cycles of copying via the polymerase chain reaction.
- steps (a) to (d) as above which further includes the steps of:
- the second method may further include the steps of:
- fragments are separated from the vector and from other fragments of the incorrect size by gel electrophoresis.
- steps (a) to (d) then (g) to (h) can be followed once, but preferably the entire cycle (c) to (d) and (g) to (h) is repeated at least once.
- a priming step in which the genes are specifically copied, is followed by a cloning step, in which the amount of genes is increased.
- the ds cDNA is derived from mRNA.
- the mRNA is preferably be isolated from lymphocytes which have been stimulated to enhance production of mRNA.
- the set of primers are preferably different from the previous step (c), so as to enhance the specificity of copying.
- the sets of primers form a nested set.
- the first set of primers may be located within the signal sequence and constant region, as described by Larrick et al., [18], and the second set of primers entirely within the variable region, as described by Orlandi et al., [19].
- the primers of step (c) include restriction sites to facilitate subsequent cloning.
- the set of primers used in step (c) should preferably include restriction sites for introduction into expression vectors.
- step (g) possible mismatches between the primers and the template strands are corrected by “nick translation”.
- step (h) the ds cDNA is preferably cleaved with restriction enzymes at sites introduced into the primers to facilitate the cloning.
- the product ds cDNA is cloned directly into an expression vector.
- the host may be prokaryotic or eukaryotic, but is preferably bacterial.
- restriction sites in the primers and in the vector, and other features of the vector will allow the expression of complete ligands, while preserving all those features of the amino acid sequence which are typical of the (methoded) ligands.
- the primers would be chosen to allow the cloning of target sequences including at least all the three CDR sequences.
- the cloning vector would then encode a signal sequence (for secretion of the ligand), and sequences encoding the N-terminal end of the first framework region, restriction sites for cloning and then the C-terminal end of the last (fourth) framework region.
- the primers would be chosen to allow the cloning of target sequences including at least the first two CDRs.
- the cloning vector could then encode signal sequence, the N-terminal end of the first framework region, restriction sites for cloning and then the C-terminal end of the third framework region, the third CDR and fourth framework region.
- Primers and cloning vectors may likewise be devised for expression of single CDRs, particularly the third CDR, as parts of complete ligands.
- the advantage of cloning repertoires of single CDRs would permit the design of a “universal” set of framework regions, incorporating desirable properties such as solubility.
- Single ligands could be expressed alone or in combination with a complementary variable domain.
- a heavy chain variable domain can be expressed either as an individual domain or, if it is expressed with a complementary light chain variable domain, as an antigen binding site.
- the two partners would be expressed in the same cell, or secreted from the same cell, and the proteins allowed to associate non-covalently to form an Fv fragment.
- the two genes encoding the complementary partners can be placed in tandem and expressed from a single vector, the vector including two sets of restriction sites.
- the genes are introduced sequentially: for example the heavy chain variable domain can be cloned first and then the light chain variable domain.
- the two genes are introduced into the vector in a single step, for example by using the polymerase chain reaction to paste together each gene with any necessary intervening sequence, as essentially described by Yon and Fried [29].
- the two partners could be also expressed as a linked protein to produce a single chain Fv fragment, using similar vectors to those described above.
- the two genes may be placed in two different vectors, for example in which one vector is a phage vector and the other is a plasmid vector.
- the cloned ds cDNA may be inserted into an expression vector already containing sequences encoding one or more constant domains to allow the vector to express Ig-type chains.
- the expression of Fab fragments would have the advantage over Fv fragments that the heavy and light chains would tend to associate through the constant domains in addition to the variable domains.
- the final expression product may be any of the modified Ig-type molecules referred to above.
- the cloned sequence may also be inserted into an expression vector so that it can be expressed as a fusion protein.
- the variable domain encoding sequence may be linked directly or via a linker sequence to a DNA sequence encoding any protein effector molecule, such as a toxin, enzyme, label or another ligand.
- the variable domain sequences may also be linked to proteins on the outer side of bacteria or phage.
- the method of this aspect of the invention may be used to produce receptors according to the invention.
- the cloning of ds cDNA directly for expression permits the rapid construction of expression libraries which can be screened for binding activities.
- the ds cDNA may comprise variable genes isolated as complete rearranged genes from the animal, or variable genes built from several different sources, for example a repertoire of unrearranged VH genes combined with a synthetic repertoire of DH and JH genes.
- repertoires of genes encoding Ig heavy chain variable domains are prepared from lymphocytes of animals immunized with an antigen.
- the screening method may take a range of formats well known in the art.
- Ig heavy chain variable domains secreted from bacteria may be screened by binding to antigen on a solid phase, and detecting the captured domains by antibodies.
- the domains may be screened by growing the bacteria in liquid culture and binding to antigen coated on the surface of ELISA plates.
- bacterial colonies or phage plaques which secrete ligands (or modified ligands, or ligand fusions with proteins) are screened for antigen binding on membranes.
- Either the ligands are bound directly to the membranes (and for example detected with labelled antigen), or captured on antigen coated membranes (and detected with reagents specific for ligands).
- the use of membranes offers great convenience in screening many clones, and such techniques are well known in the art.
- the screening method may also be greatly facilitated by making protein fusions with the ligands, for example by introducing a peptide tag which is recognized by an antibody at the N-terminal or C-terminal end of the ligand, or joining the ligand to an enzyme which catalyses the conversion of a colorless substrate to a colored product.
- the binding of antigen may be detected simply by adding substrate.
- joining of the ligand and a domain of a transcriptional activator such as the GAL4 protein of yeast, and joining of antigen to the other domain of the GAL4 protein, could form the basis for screening binding activities, as described by Fields and Song [21].
- the preparation of proteins, or even cells with multiple copies of the ligands may improve the avidity of the ligand for immobilized antigen, and hence the sensitivity of the screening method.
- the ligand may be joined to a protein subunit of a multimeric protein, to a phage coat protein or to an outer membrane protein of E. coli such as ompA or lamB.
- Such fusions to phage or bacterial proteins also offers possibilities of selecting bacteria displaying ligands with antigen binding activities.
- bacteria may be precipitated with antigen bound to a solid support, or may be subjected to affinity chromatography, or may be bound to larger cells or particles which have been coated with antigen and sorted using a fluorescence activated cell sorter (FACS).
- FACS fluorescence activated cell sorter
- the proteins or peptides fused to the ligands are preferably encoded by the vector, such that cloning of the ds cDNA repertoire creates the fusion product.
- the associated Ig heavy and light chain variable domains For example, repertoires of heavy and light chain variable genes may be cloned such that two domains are expressed together. Only some of the pairs of domains may associate, and only some of these associated pairs may bind to antigen.
- the repertoires of heavy and light chain variable domains could be cloned such that each domain is paired at random. This approach may be most suitable for isolation of associated domains in which the presence of both partners is required to form a cleft. Alternatively, to allow the binding of hapten.
- a small repertoire of light chain variable domains for example including representative members of each family of domains, may be combined with a large repertoire of heavy chain variable domains.
- a repertoire of heavy chain variable domains is screened first for antigen binding in the absence of the light chain partner, and then only those heavy chain variable domains binding to antigen are combined with the repertoire of light chain variable domains. Binding of associated heavy and light chain variable domains may be distinguished readily from binding of single domains, for example by fusing each domain to a different C-terminal peptide tag which are specifically recognized by different monoclonal antibodies.
- the hierarchical approach of first cloning heavy chain variable domains with binding activities, then cloning matching light chain variable domains may be particularly appropriate for the construction of catalytic antibodies, as the heavy chain may be screened first for substrate binding.
- a light chain variable domain would then be identified which is capable of association with the heavy chain, and “catalytic” residues such as cysteine or histidine (or prosthetic groups) would be introduced into the CDRs to stabilize the transition state or attack the substrate, as described by Baldwin and Schultz [22].
- Fab fragments are more likely to be associated than the Fv fragments, as the heavy chain variable domain is attached to a single heavy chain constant domain, and the light chain variable domain is attached to a single light chain variable domain, and the two constant domains associate together.
- the heavy and light chain variable domains are covalently linked together with a peptide, as in the single chain antibodies, or peptide sequences attached, preferably at the C-terminal end which will associate through forming cysteine bonds or through non-covalent interactions, such as the introduction of “leucine zipper” motifs.
- the Fv fragments are preferably used.
- variable domains isolated from a repertoire of variable region genes offer a way of building complete antibodies, and an alternative to hybridoma technology.
- complete antibodies may be made and should possess natural effector functions, such as complement lysis.
- This route is particularly attractive for the construction of human monoclonal antibodies, as hybridoma technology has proved difficult, and for example, although human peripheral blood lymphocytes can be immortalized with Epstein Barr virus, such hybridomas tend to secrete low affinity IgM antibodies.
- lymphocytes do not generally secrete antibodies directed against host-proteins.
- human antibodies directed against human proteins for example to human cell surface markers to treat cancers, or to histocompatibility antigens to treat auto-immune diseases.
- the construction of human antibodies built from the combinatorial repertoire of heavy and light chain variable domains may overcome this problem, as it will allow human antibodies to be built with specificities which would normally have been eliminated.
- the method also offers a new way of making bispecific antibodies.
- Antibodies with dual specificity can be made by fusing two hybridomas of different specificities, so as to make a hybrid antibody with an Fab arm of one specificity, and the other Fab arm of a second specificity.
- the yields of the bispecific antibody are low, as heavy and light chains also find the wrong partners.
- the construction of Fv fragments which are tightly associated should preferentially drive the association of the correct pairs of heavy with light chains. (It would not assist in the correct pairing of the two heavy chains with each other.)
- the improved production of bispecific antibodies would have a variety of applications in diagnosis and therapy, as is well known.
- the invention provides a species specific general oligonucleotide primer or a mixture of such primers useful for cloning variable domain encoding sequences from animals of that species.
- the method allows a single pair or pair of mixtures of species specific general primers to be used to clone any desired antibody specificity from that species. This eliminates the need to carry out any sequencing of the target sequence to be cloned and the need to design specific primers for each specificity to be recovered.
- variable genes for the expression of the variable genes directly on cloning, for the screening of the encoded domains for binding activities and for the assembly of the domains with other variable domains derived from the repertoire.
- mouse splenic ds mRNA or genomic DNA may be obtained from a hyper-immunized mouse.
- the expression vector would be used to transform a host cell, for instance a bacterial cell, to enable it to produce an Fv fragment or a Fab fragment.
- the Fv or Fab fragment would then be built into a monoclonal antibody by attaching constant domains and expressing it in mammalian cells.
- oligonucleotide primers or mixed primers were used. Their locations are marked on FIG. 1 and sequences are as follows:
- VH1FOR 5′ TGAGGAGACGGTGACCGTGGTCCCTTGGCCCCAG 3′; VH1FOR-2 5′ TGAGGAGACGGTGACCGTGGTCCCTTGGCCCC 3′; Hu1VHFOR 5′ CTTGGTGGAGGCTGAGGAGACGGTGACC 3′; Hu2VHFOR 5′ CTTGGTGGAGGCTGAGGAGACGGTGACC 3′; Hu3VHFOR 5′ CTTGGTGGATGCTGAGGAGACGGTGACC 3′; Hu4VHFOR 5′ CTTGGTGGATGCTGATGAGACGGTGACC 3′; MOJH1FOR 5′ TGAGGAGACGGTGACCGTGGTCCCTGCGCCCCCCAG 3′; MOJH2FOR 5′ TGAGGAGACGGTGACCGTGGTGCCTTGGCCCCAG 3′; MOJH3FOR 5′ TGCAGAGACGGTGACCAGTCCCTTGGCCCCAG 3′; MOJH4FOR 5′ TGAGGAGACGGTGACCGAGGTTCC
- VH1FOR is designed to anneal with the 3′ end of the sense strand of any mouse heavy chain variable domain encoding sequence. It contains a BstEII recognition site.
- VK1FOR is designed to anneal with the 3′ end of the sense strand of any mouse kappa-type light chain variable domain encoding sequence and contains a BglII recognition site.
- VH1BACK is designed to anneal with the 3′ end of the antisense strand of any mouse heavy chain variable domain and contains a PstI recognition site.
- VK1BACK is designed to anneal with the 3′ end of the antisense strand of any mouse kappa-type light chain variable domain encoding sequence and contains a PvuII recognition site.
- MAbs monoclonal antibodies
- MBr1 BW431/26 [24]
- BW494/32 BW494/32 [25]
- BW250/183 [24,26] BW704/152 [27].
- MAb MBr1 is particularly interesting in that it is known to be specific for a saccharide epitope on a human mammary carcinoma line MCF-7 [28].
- a 50 ⁇ l reaction solution which contains 10 ⁇ g mRNA, 20 pmole VH1FOR primer, 250 ⁇ M each of dATP, dTTP, dCTP and dGTP, 10 mM dithiothreitol (DTT), 100 mM Tris.HCl, 10 MM MgCl 2 and 140 mM KCl, adjusted to pH 8.3 was prepared.
- the reaction solution was heated at 70° C. for ten minutes and allowed to cool to anneal the primer to the 3′ end of the variable domain encoding sequence in the mRNA.
- To the reaction solution was then added 46 units of reverse transcriptase (Anglian Biotec) and the solution was then incubated at 42° C. for 1 hour to cause first strand cDNA synthesis.
- variable domain encoding sequences were amplified as follows.
- a 50 ⁇ l reaction solution containing 5 ⁇ l of the ds RNA/DNA hybrid-containing solution, 25 pmole each of VH1FOR and VH1BACK primers, 250 ⁇ M of dATP, dTTP, dCTP and dGTP, 67 mM Tris.HCl, 17 mM ammonium sulphate, 10 mM MgCl 2 , 200 ⁇ g/ml gelatine and 2 units Taq polymerase (Cetus) was prepared.
- the reaction solution was overlaid with paraffin oil and subjected to 25 rounds of temperature cycling using a Techne PHC-1 programmable heating block. Each cycle consisted of 1 minute and 95° C. (to denature the nucleic acids), 1 minute at 30° C. (to anneal the primers to the nucleic acids) and 2 minutes at 72° C. (to cause elongation from the primers). After the 25 cycles, the reaction solution and the oil were extracted twice with ether, once with phenol and once with phenol/CHCl3. Thereafter ds cDNA was precipitated with ethanol. The precipitated ds cDNA was then taken up in 50 ⁇ l of water and frozen.
- VK1FOR and VK1BACK primers were used in place of the VH1FOR and VH1BACK primers respectively.
- a BstEII recognition site was introduced into the vector M13-HuVHNP [31] by site directed mutagenesis [32,33] to produce the vector M13-VHPCR1 ( FIGS. 2 and 3 ).
- Each amplified heavy chain variable domain encoding sequence was digested with the restriction enzymes PstI and BstEII.
- the fragments were phenol extracted, purified on 2% low melting point agarose gels and force cloned into vector M13-VHPCR1 which had been digested with PstI and BstEII and purified on an 0.8% agarose gel.
- Clones containing the variable domain inserts were identified directly by sequencing [34] using primers based in the 3′ non-coding variable gene in the M13-VHPCR1 vector.
- variable domain encoding sequences of BW431/26 There is an internal PstI site in the heavy chain variable domain encoding sequences of BW431/26. This variable domain encoding sequence was therefore assembled in two steps. The 3′ PstI-BstEII fragment was first cloned into M13-VHPCR1, followed in a second step by the 5′ PstI fragment.
- Vector M13 mp 18 [35] was cut with PvuII and the vector backbone was blunt ligated to a synthetic HindIII-BamHI polylinker.
- Vector M13-HuVKLYS [36] was digested with HindIII and BamHI to isolate the HuVKLYS gene. This HindIII-BamHI fragment was then inserted into the HindIII-BamHI polylinker site to form a vector M13-VKPCR1 which lacks any PvuII sites in the vector backbone ( FIGS. 4 and 5 ).
- This vector was prepared in E. coli JM110 [22] to avoid dam methylation at the BclI site.
- Each amplified light chain variable domain encoding sequence was digested with PvuII and BglII.
- the fragments were phenol extracted, purified on 2% low melting point agarose gels and force cloned into vector M13-VKPCR1 which had been digested with PvuII and BclI, purified on an 0.8% agarose gel and treated with calf intestinal phosphatase.
- Clones containing the light chain variable region inserts were identified directly by sequencing [34] using primers based in the 3′ non-coding region of the variable domain in the M13-VKPCR1 vector.
- nucleotide sequences of the MBr1 heavy and light chain variable domains are shown in FIG. 6 with part of the flanking regions of the M13-VHPCR1 and M13-VKPCR1 vectors.
- the HindIII-BamHI fragment carrying the MBr1 heavy chain variable domain encoding sequence in M13-VHPCR1 was recloned into a pSV-gpt vector with human ⁇ 1 constant regions [37] ( FIG. 7 ).
- the MBr1 light chain variable domain encoding sequence in M13-VKPCR1 was recloned as a HindIII-BamHI fragment into a pSV vector, PSV-hyg-HuCK with a hygromycin resistance marker and a human kappa constant domain ( FIG. 8 ).
- the assembly of the genes is summarized in FIG. 9 .
- the vectors thus produced were linearized with PvuI (in the case of the pSV-hygro vectors the PvuI digest is only partial) and cotransfected into the non-secreting mouse myeloma line NSO [38] by electroporation [39].
- PvuI in the case of the pSV-hygro vectors the PvuI digest is only partial
- NSO non-secreting mouse myeloma line NSO
- electroporation electroporation [39].
- One day after cotransfection cells were selected in 0.3 ⁇ g/ml mycophenolic acid (MPA) and after seven days in 1 ⁇ g/ml MPA. After 14 days, four wells, each containing one or two major colonies, were screened by incorporation of 14 C-lysine [40] and the secreted antibody detected after precipitation with protein-A SepharoseTM (Pharmacia) on SDS-PAGE [41].
- the gels were stained, fixed, soaked in
- the chimeric antibody in the supernatant like the parent mouse MBr1 antibody, was found to bind to MCF-7 cells but not the HT-29 cells, thus showing that the specificity had been properly cloned and expressed.
- the DNA from the mouse spleen was prepared in one of two ways (although other ways can be used).
- Method 1 A mouse spleen was cut into two pieces and each piece was put into a standard Eppendorf tube with 200 ⁇ l of PBS. The tip of a 1 ml glass pipette was closed and rounded in the blue flame of a Bunsen burner. The pipette was used to squash the spleen piece in each tube. The cells thus produced were transferred to a fresh Eppendorf tube and the method was repeated three times until the connective tissue of the spleen appeared white. Any connective tissue which has been transferred with the cells was removed using a drawn-out Pasteur pipette. The cells were then washed in PBS and distributed into four tubes.
- mice spleen cells were then sedimented by a 2 minute spin in a Microcentaur centrifuge at low speed setting. All the supernatant was aspirated with a drawn out Pasteur pipette. If desired, at this point the cell sample can be frozen and stored at ⁇ 20° C.
- the supernatant was transferred to a new tube and to this was added 125 ⁇ l 5M NaCl and 30 ⁇ l 1M MOPS adjusted to pH 7.0.
- the DNA in the supernatant was absorbed on a Quiagen 5 tip and purified following the manufacturer's instructions for lambda DNA. After isopropanol precipitation, the DNA was resuspended in 500 ⁇ l water.
- Method 2 This method is based on the technique described in Maniatis et al. [30].
- a mouse spleen was cut into very fine pieces and put into a 2 ml glass homogenizer. The cells were then freed from the tissue by several slow up and down strokes with the piston.
- the cell suspension was made in 500 ⁇ l phosphate buffered saline (PBS) and transferred to an Eppendorf tube. The cells were then spun for 2 min at low speed in a Microcentaur centrifuge. This results in a visible separation of white and red cells. The white cells, sedimenting slower, form a layer on top of the red cells. The supernatant was carefully removed and spun to ensure that all the white cells had sedimented.
- the layer of white cells was resuspended in two portions of 500 ⁇ l PBS and transferred to another tube.
- the white cells were precipitated by spinning in the Microcentaur centrifuge at low speed for one minute. The cells were washed a further two times with 500 ⁇ l PBS, and were finally resuspended in 200 ⁇ l PBS. The white cells were added to 2.5 ml 25 mM EDTA and 10 mM Tris.Cl, pH 7.4, and vortexed slowly. While vortexing 25 ⁇ l 20% SDS was added. The cells lysed immediately and the solution became viscous and clear. 100 ⁇ l of 20 mg/ml proteinase K was added and incubated one to three hours at 50° C.
- the sample was extracted with an equal volume of phenol and the same volume of chloroform, and vortexed. After centrifuging, the aqueous phase was removed and 1/10 volume 3M ammonium acetate was added. This was overlaid with three volumes of cold ethanol and the tube rocked carefully until the DNA strands became visible.
- the DNA was spooled out with a Pasteur pipette, the ethanol allowed to drip off, and the DNA transferred to 1 ml of 10 mM Tris.Cl pH 7.4, 0.1 mM EDTA in an Eppendorf tube. The DNA was allowed to dissolve in the cold overnight on a roller.
- the DNA solution was diluted 1/10 in water and boiled for 5 min prior to using the polymerase chain reaction (PCR).
- PCR polymerase chain reaction
- typically 50-200 ng of DNA were used.
- the heavy and light chain variable domain encoding sequences in the genomic DNA isolated from the human PBL or the mouse spleen cells was then amplified and cloned using the general protocol described in the first two paragraphs of the section headed “Amplification from RNA/DNA Hybrid” in Example 1, except that during the annealing part of each cycle, the temperature was held at 65° C. and that 30 cycles were used. Furthermore, to minimize the annealing between the 3′ ends of the two primers, the sample was first heated to 95° C., then annealed at 65° C., and only then was the Taq polymerase added. At the end of the 30 cycles, the reaction mixture was held at 60° C. for five minutes to ensure that complete elongation and renaturation of the amplified fragments had taken place.
- VH1FOR and VH1BACK The primers used to amplify the mouse spleen genomic DNA were VH1FOR and VH1BACK, for the heavy chain variable domain and VK2FOR and VK1BACK, for the light chain variable domain. (VK2FOR only differs from VK1FOR in that it has an extra C residue on the 5′ end.)
- VH1FOR Likewise mixtures of VH1FOR, MOJH1FOR, MOJH2FOR, MOJH3FOR and MOJH4FOR were used as forward primers and mixtures of VH1BACK, MOVHIBACK, MOVHIIABACK, MOVHIIBBACK, MOVHIIIBACK were used as backward primers for amplification of VH genes.
- All these heavy chain FOR primers referred to above contain a BstEII site and all the BACK primers referred to above contain a PstI site. These light chain FOR and BACK primers referred to above all contain BglII and PvuII sites respectively.
- Light chain primers VK3FOR and VK2BACK were also devised which utilized different restriction sites, SacI and XhoI.
- the preferred amplification conditions for mouse VH genes are as follows: the sample was made in a volume of 50-100 ⁇ l, 50-100 ng of DNA, VH1FOR-2 and VH1BACK primers (25 pmole of each), 250 ⁇ M of each deoxynucleotide triphosphate, 10 mM Tris.HCl, pH 8.8, 50 mM KCl, 1.5 mM MgCl 2 , and 100 ⁇ g/ml gelatine. The sample was overlaid with paraffin oil, heated to 95° C. for 2 min, 65° C.
- the preferred amplification conditions for mouse V ⁇ genes from genomic DNA are as follows: the sample treated as above except with V ⁇ primers, for example VK3FOR and VK2BACK, and using a cycle of 94° C. for one minute, 60° C. for one minute and 72° C. for one minute.
- V ⁇ primers for example VK3FOR and VK2BACK
- the conditions which were devised for genomic DNA are also suitable for amplification from the cDNA derived from mRNA from mouse spleen or mouse hybridoma.
- the reaction mixture was then extracted twice with 40 ⁇ l of water-saturated diethyl ether. This was followed by a standard phenol extraction and ethanol precipitation as described in Example 1.
- the DNA pellet was then dissolved in 100 ⁇ l 10 mM Tris.Cl, 0.1 mM EDTA.
- Each reaction mixture containing a light chain variable domain encoding sequence was digested with SacI and XhoI (or with PvuII and BglII) to enable it to be ligated into a suitable expression vector.
- Each reaction mixture containing a heavy chain variable domain encoding sequence was digested with PstI and BstEII for the same purpose.
- the heavy chain variable genes isolated as above from a mouse hyper-immunized with lysozyme were cloned into M13VHPCR1 vector and sequenced.
- the complete sequences of 48 VH gene clones were determined ( FIGS. 10 a - 10 b ). All but two of the mouse VH gene families were represented, with frequencies of: VA (1), IIIC (1), IIIB (8), IIIA (3), IIB (17), IIA (2), IB (12), IA (4).
- the D segments could be assigned to families SP2 (14), FL16 (11) and Q52 (5), and in 38 clones the JH minigenes to families JH1 (3), JH2 (7), JH3 (14) and JH4 (14).
- the different sequences of CDR3 marked out each of the 48 clones as unique. Nine pseudogenes and 16 unproductive rearrangements were identified. Of the clones sequenced, 27 have open reading frames.
- the method is capable of generating a diverse repertoire of heavy chain variable genes from mouse spleen DNA.
- Method 1 20 ml of heparinized human blood from a healthy volunteer was diluted with an equal volume of phosphate buffered saline (PBS) and distributed equally into 50 ml Falcon tubes. The blood was then underlayed with 15 ml Ficoll Hypaque (Pharmacia 10-A-001-07). To separate the lymphocytes from the red blood cells, the tubes were spun for 10 minutes at 1800 rpm at room temperature in an IEC Centra 3E table centrifuge. The peripheral blood lymphocytes (PBL) were then collected from the interphase by aspiration with a Pasteur pipette. The cells were diluted with an equal volume of PBS and spun again at 1500 rpm for 15 minutes. The supernatant was aspirated, the cell pellet was resuspended in 1 ml PBS and the cells were distributed into two Eppendorf tubes.
- PBS phosphate buffered saline
- Method 2 40 ml human blood from a patient with HIV in the pre-AIDS condition was layered on Ficoll to separate the white cells (see Method 1 above). The white cells were then incubated in tissue culture medium for 4-5 days. On day 3, they were infected with Epstein Barr virus. The cells were pelleted (approx 2 ⁇ 10 7 cells) and washed in PBS.
- the cells were pelleted again and lysed with 7 ml 5M guanidine isothiocyanate, 50 mM Tris, 10 mM EDTA, 0.1 mM dithiothreitol.
- the cells were vortexed vigorously and 7 volumes of 4M LiCl added.
- the mixture was incubated at 4° C. for 15-20 hrs.
- the suspension was spun and the supernatant resuspended in 3M LiCl and centrifuged again.
- the pellet was dissolved in 2 ml 0.1% SDS, 10 mM Tris HCl and 1 mM EDTA.
- the suspension was frozen at ⁇ 20° C., and thawed by vortexing for 20 s every 10 min for 45 min.
- RNA was precipitated by adding 1/10 volume 3M sodium acetate and 2 vol ethanol and leaving overnight at ⁇ 20° C. The pellet was suspended in 0.2 ml water and reprecipitated with ethanol. Aliquots for cDNA synthesis were taken from the ethanol precipitate which had been vortexed to create a fine suspension.
- the back primers for the amplification of human DNA were designed to match the available human heavy and light chain sequences, in which the different families have slightly different nucleotide sequences at the 5′ end.
- the primers Hu2VHIBACK, HuVHIIBACK, Hu2VHIIIBACK and HuVH1VBACK were designed as back primers, and HUJH1FOR, HUJH2FOR and HUJH4FOR as forward primers based entirely in the variable gene.
- Another set of forward primers Hu1VHFOR, Hu2VHFOR, Hu3VHFOR, and Hu4VHFOR was also used, which were designed to match the human J-regions and the 5′ end of the constant regions of different human isotypes.
- the amplified DNA from the separate primings was then pooled, digested with restriction enzymes PstI and BstEII as above, and then cloned into the vector M13VHPCR1 for sequencing.
- the sequences reveal a diverse repertoire ( FIG. 11 ) at this stage of the disease.
- HuJK1FOR, HUJK3FOR; HUJK4FOR and HUJK5FOR were used as forward primers and VK1BACK as back primer. Using these primers it was possible to see a band of amplified ds cDNA of the correct size by gel electrophoresis.
- Human peripheral blood lymphocytes of a patient with non-Hodgkins lymphoma were prepared as in Example 3 (Method 1).
- the genomic DNA was prepared from the PBL using the technique described in Example 2 (Method 2).
- the VH region in the isolated genomic DNA was then amplified and cloned using the general protocol described in the first two paragraphs of the section headed “Amplification from RNA/DNA hybrid” in Example 1 above, except that during the annealing part of each cycle, the temperature was held at 55° C. and that 30 cycles were used. At the end of the 30 cycles, the reaction mixture was held at 60° C. for five minutes to ensure that complete elongation and renaturation of the amplified fragments had taken place.
- the forward primer used was HuHep1FOR, which contains a SacI site. This primer is designed to anneal to the 3′ end of the unrearranged human VH region gene, and in particular includes a sequence complementary to the last three codons in the VH region gene and nine nucleotides downstream of these three codons.
- HuOcta1BACK, HuOcta2BACK and HuOcta3BACK was used as the back primer. These primers anneal to a sequence in the promoter region of the genomic DNA VH gene (see FIG. 1 ). 5 ⁇ l of the amplified DNA was checked on 2% agarose gels in TBE buffer and stained with ethidium bromide. A double band was seen of about 620 nucleotides which corresponds to the size expected for the unrearranged VH gene. The ds cDNA was digested with SacI and cloned into an M13 vector for sequencing. Although there are some sequences which are identical, a range of different unrearranged human VH genes were identified ( FIG. 12 ).
- VHLYS heavy chain variable domain
- D1.3 anti-lysozyme
- the heavy chain variable domain (VHLYS) of the D1.3 (anti-lysozyme) antibody was cloned into a vector similar to that described previously [42] but under the control of the lac z promoter, such that the VHLYS domain is attached to a pelB leader sequence for export into the periplasm.
- the vector was constructed by synthesis of the pelB leader sequence [43], using overlapping oligonucleotides, and cloning into a pUC 19 vector [35].
- the VHLYS domain of the D1.3 antibody was derived from a cDNA clone [44] and the construct (PSW1) sequenced ( FIG. 13 ).
- VKLYS light chain variable region
- the colonies were inoculated into 50 ml 2 ⁇ TY (with 1% glucose and 100 ⁇ g/ml ampicillin) and grown in flasks at 37° C. with shaking for 12-16 hr.
- the cells were centrifuged, the pellet washed twice with 50 mM sodium chloride, resuspended in 2 ⁇ TY medium containing 100 ⁇ g/ml ampicillin and the inducer IPTG (1 mM) and grown for a further 30 hrs at 37° C.
- the cells were centrifuged and the supernatant was passed through a Nalgene filter (0.45 ⁇ m) and then down a 1-5 ml lysozyme-Sepharose® affinity column (Pharmacia Fine Chemicals, Inc.).
- the column was derived by coupling lysozyme at 10 mg/ml to CNBr activated Sepharose.
- the column was first washed with phosphate buffered saline (PBS), then with 50 mM diethylamine to elute the VHLYS domain (from pSW1) or VHLYS in association with VKLYS (from pSW2).
- PBS phosphate buffered saline
- VHLYS and VKLYS domains were identified by SDS polyacrylamide electrophoresis as the correct size.
- N-terminal sequence determination of VHLYS and VKLYS isolated from a polyacrylamide gel showed that the signal peptide had been produced correctly.
- both the Fv fragment and the VHLYS domains are able to bind to the lysozyme affinity column, suggesting that both retain at least some of the affinity of the original antibody.
- VHLYS domain was compared by FPLC with that of the Fv fragment on Superose 12. This indicates that the VHLYS domain is a monomer.
- the binding of the VHLYS and Fv fragment to lysozyme was checked by ELISA, and equilibrium and rapid reaction studies were carried out using fluorescence quench.
- the ELISA for lysozyme binding was undertaken as follows:
- VHLYS or Fv fragment VHLYS associated with VKLYS
- the reaction was stopped by adding 0.05% sodium azide in 50 mM citric acid pH 4.3.
- ELISA plates were read in a Titertek Multiscan plate reader. Supernatant from the induced bacterial cultures of both pSW1 (VHLYS domain) or pSW2 (Fv fragment) was found to bind to lysozyme in the ELISA.
- the purified VHLYS and Fv fragments were titrated with lysozyme using fluorescence quench (Perkin Elmer LS5B Luminescence Spectrometer) to measure the stoichiometry of binding and the affinity constant for lysozyme [48,49].
- fluorescence quench Perkin Elmer LS5B Luminescence Spectrometer
- the titration of the Fv fragment at a concentration of 30 nM indicates a dissociation constant of 2.8 nM using a Scatchard analysis.
- VHLYS was titrated with lysozyme as above using fluorescence quench.
- the on-rates for VHLYS and Fv fragments with lysozyme were determined by stopped-flow analysis (HI Tech Stop Flow SHU machine) under pseudo-first order conditions with the fragment at a ten fold higher concentration than lysozyme [50].
- the concentration of lysozyme binding sites was first measured by titration with lysozyme using fluorescence quench as above. The on rates were calculated per mole of binding site (rather than amount of VHLYS protein).
- the on-rate for the Fv fragment was found to be 2.2 ⁇ 10 6 M ⁇ 1 s ⁇ 1 at 25° C.
- the on-rate for the VHLYS fragment found to be 3.8 ⁇ 10 6 M ⁇ 1 s ⁇ 1 and the off-rate 0.075 s ⁇ 1 at 20° C.
- the calculated affinity constant is 19 nM.
- the VHLYS binds to lysozyme with a dissociation constant of about 19 nM, compared with that of the Fv of 3 nM.
- a mouse was immunized with hen egg white lysozyme (100 ⁇ g i.p. day 1 in complete Freunds adjuvant), after 14 days immunized i.p. again with 100 ⁇ g lysozyme with incomplete Freunds adjuvant, and on day 35 i.v. with 50 ⁇ g lysozyme in saline. On day 39, spleen was harvested. A second mouse was immunized with keyhole limpet hemocyanin (KLH) in a similar way. The DNA was prepared from the spleen according to Example 2 (Method 2). The VH genes were amplified according to the preferred method in Example 2.
- KLH keyhole limpet hemocyanin
- Human peripheral blood lymphocytes from a patient infected with HIV were prepared as in Example 3 (Method 2) and mRNA prepared.
- the VH genes were amplified according to the method described in Example 3, using primers designed for human VH gene families.
- the reaction mixture and oil were extracted twice with ether, once with phenol and once with phenol/CHCl 3 .
- the double stranded DNA was then taken up in 50 ⁇ l of water and frozen. 5 ⁇ l was digested with PstI and BstEII (encoded within the amplification primers) and loaded on an agarose gel for electrophoresis. The band of amplified DNA at about 350 bp was extracted.
- the repertoire of amplified heavy chain variable domains was then cloned directly into the expression vector pSW1HPOLYMYC.
- This vector is derived from pSW1 except that the VHLYS gene has been removed and replaced by a polylinker restriction site.
- a sequence encoding a peptide tag was inserted ( FIG. 15 ). Colonies were toothpicked into 1 ml cultures. After induction (see Example 5 for details), 10 ⁇ l of the supernatant from fourteen 1 ml cultures was loaded on SDS-PAGE gels and the proteins transferred electrophoretically to nitrocellulose. The blot was probed with antibody 9E10 directed against the peptide tag.
- the probing was undertaken as follows.
- the nitrocellulose filter was incubated in 3% bovine serum albumin (BSA)/TBS buffer for 20 min (10 ⁇ TBS buffer is 100 mM Tris.HCl, pH 7.4, 9% w/v NaCl).
- BSA bovine serum albumin
- the filter was incubated in a suitable dilution of antibody 9E10 (about 1/500) in 3% BSA/TBS for 1-4 hrs. After three washes in TBS (100 ml per wash, each wash for 10 min), the filter was incubated with 1:500 dilution of anti-mouse antibody (peroxidase conjugated anti-mouse Ig (Dakopats)) in 3% BSA/TBS for 1-2 hrs.
- anti-mouse antibody peroxidase conjugated anti-mouse Ig (Dakopats)
- Colonies were then toothpicked individually into wells of an ELISA plate (200 ⁇ l) for growth and induction. They were assayed for lysozyme binding with the 9E10 antibody (as in Examples 5 and 7). Wells with lysozyme-binding activity were identified. Two positive wells (of 200) were identified from the amplified mouse spleen DNA and one well from the human cDNA. The heavy chain variable domains were purified on a column of lysozyme-Sepharose. The affinity for lysozyme of the clones was estimated by fluorescence quench titration as >50 nM.
- VH3 and VH8 The affinities of the two clones (VH3 and VH8) derived from the mouse genes were also estimated by stop flow analysis (ratio of k off /k on ) as 12 nM and 27 nM respectively. Thus both these clones have a comparable affinity to the VHLYS domain.
- the encoded amino acid sequences of VH3 and VH8 are given in FIG. 16 , and that of the human variable domain in FIG. 17 .
- a library of VH domains made from the mouse immunized with lysozyme was screened for both lysozyme and keyhole limpet hemocyanin (KLH) binding activities. Two thousand colonies were toothpicked in groups of five into wells of ELISA plates, and the supernatants tested for binding to lysozyme coated plates and separately to KLH coated plates. Twenty one supernatants were shown to have lysozyme binding activities and two to have KLH binding activities.
- a second expression library, prepared from a mouse immunized with KLH was screened as above. Fourteen supernatants had KLH binding activities and a single supernatant had lysozyme binding activity.
- a single rearranged VH gene it may be possible to derive entirely new antigen binding activities by extensively mutating each of the CDRs.
- the mutagenesis might be entirely random, or be derived from pre-existing repertoires of CDRs.
- a repertoire of CDR3s might be prepared as in the preceding examples by using “universal” primers based in the flanking sequences, and likewise repertoires of the other CDRs (singly or in combination).
- the CDR repertoires could be stitched into place in the flanking framework regions by a variety of recombinant DNA techniques.
- CDR3 appears to be the most promising region for mutagenesis as CDR3 is more variable in size and sequence than CDRs 1 and 2. This region would be expected to make a major contribution to antigen binding.
- the heavy chain variable region (VHLYS) of the anti-lysozyme antibody D1.3 is known to make several important contacts in the CDR3 region.
- the source of the heavy chain variable domain was an M113 vector containing the VHLYS gene.
- the body of the sequence encoding the variable region was amplified using the polymerase chain reaction (PCR) with the mutagenic primer VHMUT1 based in CDR3 and the M13 primer which is based in the M13 vector backbone.
- the mutagenic primer hypermutates the central four residues of CDR3 (Arg-Asp-Tyr-Arg).
- the PCR was carried out for 25 cycles on a Techne PHC-1 programmable heat block using 100 ng single stranded M13 mp19SWO template, with 25 pmol of VHMUT1 and the M13 primer, 0.5 mM each dNTP, 67 mM Tris.HCl, pH 8.8, 10 mM MgCl 2 , 17 mM (NH 4 ) 2 SO 4 , 200 ⁇ g/ml gelatine and 2.5 units Taq polymerase in a final volume of 50 ⁇ l.
- the temperature regime was 95° C. for 1.5 min, 25° C. for 1.5 min and 72° C. for 3 min (However a range of PCR conditions could be used).
- the reaction products were extracted with phenol/chloroform, precipitated with ethanol and resuspended in 10 mM Tris. HCl and 0.1 mM EDTA, pH 8.0.
- the products from the PCR were digested with PstI and BstEII and purified on a 1.5% LGT agarose gel in Tris acetate buffer using Geneclean® (Bio 101, LaJolla).
- the gel purified band was ligated into pSW2HPOLY ( FIG. 19 ).
- the vector was first digested with BstEII and PstI and treated with calf-intestinal phosphatase. Aliquots of the reaction mix were used to transform E. coli BMH 71-18 to ampicillin resistance. Colonies were selected on ampicillin (100 ⁇ g/ml) rich plates containing glucose at 0.8% w/v.
- Colonies resulting from transfection were picked in pools of five into two 96 well Corning microtitre plates, containing 200 ⁇ l 2 ⁇ TY medium and 100 ⁇ l TY medium, 100 ⁇ g/ml ampicillin and 1% glucose. The colonies were grown for 24 hours at 37° C. and then cells were washed twice in 200 ⁇ l 50 mM NaCl, pelleting the cells in an IEC Centra-3 bench top centrifuge with microtitre plate head fitting. Plates were spun at 2,500 rpm for 10 min at room temperature. Cells were resuspended in 200 ⁇ l 2 ⁇ TY, 100 ⁇ g/ml ampicillin and 1 mM IPTG (Sigma) to induce expression, and grown for a further 24 hr.
- plasmids were prepared and the VKLYS gene excised by cutting with EcoRI and religating. Thus the plasmids should only direct the expression of the VHLYS mutants. 1.5 ml cultures were grown and induced for expression as above. The cells were spun down and supernatant shown to bind lysozyme as above. (Alternatively the amplified mutant VKLYS genes could have been cloned directly into the pSW1HPOLY vector for expression of the mutant activities in the absence of VKLYS.)
- An ELISA method was devised in which the activities of bacterial supernatants for binding of lysozyme (or KLH) were compared.
- a vector was devised for tagging of the VH domains at its C-terminal region with a peptide from the c-myc protein which is recognized by a monoclonal antibody 9E10.
- the vector was derived from pSW1 by a BstEII and SmaI double digest, and ligation of an oligonucleotide duplex made from
- VHLYSMYC protein domain expressed after induction was shown to bind to lysozyme and to the 9E10 antibody by ELISA as follows:
- test solution was discarded, and the wells washed out with PBS (3 washes).
- 100 ⁇ l of 4 ⁇ g/ml purified 9E10 antibody in 2% Sainsbury's instant dried skimmed milk powder in PBS was added, and incubated at 37° C. for 2 hrs;
- reaction was stopped by adding 0.05% sodium azide in 50 mM citric acid, pH 4.3.
- ELISA plates were read in an Titertek Multiscan plate reader.
- VHLYSMUT59 To check the affinity of the VHLYSMUT59 domain directly, the clone was grown at the 1 L scale and 200-300 ⁇ g purified on lysozyme-Sepharose as in Example 5. By fluorescence quench titration of samples of VHLYS and VHLYSMUT59, the number of binding sites for lysozyme were determined. The samples of VHLYS and VHLYSMUT59 were then compared in the competition ELISA with VHLYSMYC over two orders of magnitude. In the competition assay each microtitre well contained a constant amount of VHLYSMYC (approximately 0.6 ⁇ g VHLYSMYC).
- VHLYS or VHLYSMUT59 Varying amounts of VHLYS or VHLYSMUT59 (3.8 ⁇ M in lysozyme binding sites) were added (0.166-25 ⁇ l). The final volume and buffer concentration in all wells was constant. 9E10 (anti-myc) antibody was used to quantitate bound VHLYSMYC in each assay well. The % inhibition of VHLYSMYC binding was calculated for each addition of VHLYS or VHLYSMUT59, after subtraction of background binding. Assays were carried out in duplicate. The results indicate that VHLYSMUT59 has a higher affinity for lysozyme than VHLYS.
- VHLYSMUT59 gene was sequenced (after recloning into M13) and shown to be identical to the VHLYS gene except for the central residues of CDR3 (Arg-Asp-Tyr-Arg). These were replaced by Thr-Gln-Arg-Pro: (encoded by ACACAAAGGCCA).
- a library of 2000 mutant VH clones was screened for lysozyme and also for KLH binding (toothpicking 5 colonies per well as described in Example 6).
- Nineteen supernatants were identified with lysozyme binding activities and four with KLH binding activities. This indicates that new specificities and improved affinities can be derived by making a random repertoire of CDR3.
- Two copies of the cloned heavy chain variable gene of the D1.3 antibody were linked by a nucleotide sequence encoding a flexible linker Gly-Gly-Gly-Ala-Pro-Ala-Ala-Ala-Pro-Ala-Gly-Gly-Gly- (by several steps of cutting, pasting and site directed mutagenesis) to yield the plasmid pSW3 ( FIG. 20 ).
- the expression was driven by a lacZ promoter and the protein was secreted into the periplasm via a pelB leader sequence (as described in Example 5 for expression of pSW1 and pSW2).
- the protein could be purified to homogeneity on a lysozyme affinity column.
- a cysteine residue was introduced at the C-terminus of the VHLYS domain in the vector pSW2.
- the cysteine was introduced by cleavage of the vector with the restriction enzymes BstI and SmaI (which excises the C-terminal portion of the J segment) and ligation of a short oligonucleotide duplex
- FIGS. 21 a - 21 c there is shown the sequence of a fusion of a VH domain with alkaline phosphatase.
- the alkaline phosphatase gene was cloned from a plasmid carrying the E. coli alkaline phosphatase gene in a plasmid pEK48 [51] using the polymerase chain reaction. The gene was amplified with the primers
- FIGS. 21 a - 21 c The construction ( FIGS. 21 a - 21 c ) was expressed in E.
- Example 5 coli strain BMH71-18 as in Example 5 and screened for phosphatase activity using 1 mg/ml p-nitrophenylphosphate as substrate in 10 mM diethanolamine and 0.5 mM MgCl 2 , pH 9.5) and also on SDS polyacrylamide gels which had been Western blotted (detecting with anti-idiotypic antiserum). No evidence was found for the secretion of the linked VHLYS-alkaline phosphatase as detected by Western blots (see Example 5), or for secretion of phosphatase activity.
- linker sequences could then be introduced at the BstEII site to improve the spacing between the two domains.
- V ⁇ genes A repertoire of V ⁇ genes was derived by PCR using primers as described in Example 2 from DNA prepared from mouse spleen and also from mouse spleen mRNA using the primers VK3FOR and VK2BACK and a cycle of 94° C. for 1 min, 60° C. for 1 min, 72° C. for 2 min.
- the PCR amplified DNA was fractionated on the agarose gel, the band excised and cloned into a vector which carries the VHLYS domain (from the D 1.3 antibody), and a cloning site (SacI and XhoI) for cloning of the light chain variable domains with a myc tail (pSW1VHLYS-VKPOLYMYC, FIG. 22 ).
- Clones were screened for lysozyme binding activities as described in Examples 5 and 7 via the myc tag on the light chain variable domain, as this should permit the following kinds of V ⁇ domains to be identified:
- VHLYS domain was replaced by the heavy chain variable domain VH3 which had been isolated from the repertoire (see Example 6), and then the V ⁇ domains cloned into the vector. (Note that the VH3 domain has an internal SacI site and this was first removed to allow the cloning of the V ⁇ repertoire as SacI-XhoI fragments.)
- the present invention enables the cloning, amplification and expression of heavy and light chain variable domain encoding sequences in a much more simple manner than was previously possible. It also shows that isolated variable domains or such domains linked to effector molecules are unexpectedly useful.
Abstract
Description
- This is a continuation of application Ser. No. 10/290,233, filed Nov. 8, 2002 (allowed), which is a continuation of application Ser. No. 09/722,364, filed Nov. 28, 2000 (now U.S. Pat. No. 6,545,142), which is a continuation of application Ser. No. 08/470,031, filed Jun. 6, 1995 (now U.S. Pat. No. 6,248,516), which is a divisional of application Ser. No. 08/332,046, filed Nov. 1, 1994 (now abandoned), which is a continuation of application Ser. No. 07/796,805, filed Nov. 25, 1991 (now abandoned), which is a divisional of application Ser. No. 07/580,374, filed Sep. 11, 1990 (now abandoned), which is a continuation of PCT Application No. PCT/GB89/01344, filed Nov. 13, 1989, the entire contents of each of which is hereby incorporated by reference in this application.
- The present invention relates to single domain ligands derived from molecules in the immunoglobulin (Ig) superfamily, receptors comprising at least one such ligand, methods for cloning, amplifying and expressing DNA sequences encoding such ligands, methods for the use of said DNA sequences in the production of Ig-type molecules and said ligands or receptors, and the use of said ligands or receptors in therapy, diagnosis or catalysis.
- The present invention is now described, by way of example only, with reference to the accompanying drawings.
-
FIG. 1 shows a schematic representation of the unrearranged and rearranged heavy and light chain variable genes and the location of the primers. -
FIG. 2 shows a schematic representation of the M13-VHPCR1 vector and a cloning scheme for amplified heavy chain variable domains. -
FIG. 3 shows the sequence of the Ig variable region derived sequences in M13-VHPCR1. -
FIG. 4 shows a schematic representation of the M13-VKPCR1 vector and a cloning scheme for light chain variable domains. -
FIG. 5 shows the sequence of the Ig variable region derived sequences in M13-VKPCR1. -
FIG. 6 shows the nucleotide sequences of the heavy and light chain variable domain encoding sequences of MAb MBr1. -
FIG. 7 shows a schematic representation of the pSV-gpt vector (also known as α-Lys 30) which contains a variable region cloned as a HindIII-BamHI fragment, which is excised on introducing the new variable region. The gene for human IgG1 has also been engineered to remove a BamHI site, such that the BamHI site in the vector is unique. -
FIG. 8 shows a schematic representation of the pSV-hygro vector (also known as α-Lys 17). It is derived from pSV gpt vector with the gene encoding mycophenolic acid replaced by a gene coding for hygromycin resistance. The construct contains a variable gene cloned as a HindIII-BamHI fragment which is excised on introducing the new variable region. The gene for human Cκ has also been engineered to remove a BamHI site, such that the BamHI site in the vector is unique. -
FIG. 9 shows the assembly of the mouse: human MBr1 chimeric antibody. -
FIGS. 10 a-10 b shows encoded amino acid sequences of 48 mouse rearranged VH genes. -
FIG. 11 shows encoded amino acid sequences of human rearranged VH genes. -
FIG. 12 shows encoded amino acid sequences of unrearranged human VH genes. -
FIG. 13 shows the sequence of part of the plasmid pSW1: essentially the sequence of a pectate lyase leader linked to VHLYS in pSW1 and cloned as an SphI-EcoRI fragment into pUC19 and the translation of the open reading frame encoding the pectate lyase leader-VHLYS polypeptide being shown. -
FIGS. 14 a-14 b shows the sequence of part of the plasmid pSW2: essentially the sequence of a pectate lyase leader linked to VHLYS and to VKLYS, and cloned as an SphI-EcoRI-EcoRI fragment into pUC19 and the translation of open reading frames encoding the pectate lyase leader-VHLYS and pectate lyase leader-VKLYS polypeptides being shown. -
FIG. 15 shows the sequence of part of the plasmid pSW1HPOLYMYC which is based on pSW1 and in which a polylinker sequence has replaced the variable domain of VHLYS, and acts as a cloning site for amplified VH genes, and a peptide tag is introduced at the C-terminal end. -
FIG. 16 shows the encoded amino acid sequences of two VH domains derived from mouse spleen and having lysozyme binding activity, and compared with the VH domain of the D1,3 antibody. The arrows mark the points of difference between the two VH domains. -
FIG. 17 shows the encoded amino acid sequence of a VH domain derived from human peripheral blood lymphocytes and having lysozyme binding activity. -
FIG. 18 shows a scheme for generating and cloning mutants of the VHLYS gene, which is compared with the scheme for cloning natural repertoires of VH genes. -
FIG. 19 shows the sequence of part of the vector pSW2HPOLY. -
FIG. 20 shows the sequence of part of the vector pSW3 which encodes the two linked VHLYS domains. -
FIGS. 21 a-21 c shows the sequence of the VHLYS domain and pelB leader sequence fused to the alkaline phosphatase gene. -
FIG. 22 shows the sequence of the vector pSW1VHLYS-VKPOLYMYC for expression of a repertoire of Vκ light chain variable domains in association with the VHLYS domain. -
FIG. 23 shows the sequence of VH domain which is secreted at high levels from E. coli. The differences with VHLYS domain are marked. - The present invention relates to single domain ligands derived from molecules in the immunoglobulin (Ig) superfamily, receptors comprising at least one such ligand, methods for cloning, amplifying and expressing DNA sequences encoding such ligands, methods for the use of said DNA sequences in the production of Ig-type molecules and said ligands or receptors, and the use of said ligands or receptors in therapy, diagnosis or catalysis.
- A list of references is appended to the end of the description. The documents listed therein are referred to in the description by number, which is given in square brackets.
- The Ig superfamily includes not only the Igs themselves but also such molecules as receptors on lymphoid cells such as T lymphocytes. Immunoglobulins comprise at least one heavy and one light chain covalently bonded together. Each chain is divided into a number of domains. At the N-terminal end of each chain is a variable domain. The variable domains on the heavy and light chains fit together to form a binding site designed to receive a particular target molecule. In the case of Igs, the target molecules are antigens. T-cell receptors have two chains of equal size, the α and β chains, each consisting of two domains. At the N-terminal end of each chain is a variable domain and the variable domains on the α and β chains are believed to fit together to form a binding site for target molecules, in this case peptides presented by a histocompatibility antigen. The variable domains are so called because their amino acid sequences vary particularly from one molecule to another. This variation in sequence enables the molecules to recognize an extremely wide variety of target molecules.
- Much research has been carried out on Ig molecules to determine how the variable domains are produced. It has been shown that each variable domain comprises a number of areas of relatively conserved sequence and three areas of hypervariable sequence. The three hypervariable areas are generally known as complementarity determining regions (CDRs).
- Crystallographic studies have shown that in each variable domain of an Ig molecule the CDRs are supported on framework areas formed by the areas of conserved sequences. The three CDRs are brought together by the framework areas and, together with the CDRs on the other chain, form a pocket in which the target molecule is received.
- Since the advent of recombinant DNA technology, there has been much interest in the use of such technology to clone and express Ig molecules and derivatives thereof. This interest is reflected in the numbers of patent applications and other publications on the subject.
- The earliest work on the cloning and expression of full Igs in the patent literature is EP-
A-0 120 694 (Boss). The Boss application also relates to the cloning and expression of chimeric antibodies. Chimeric antibodies are Ig-type molecules in which the variable domains from one Ig are fused to constant domains from another Ig. Usually, the variable domains are derived from an Ig from one species (often a mouse Ig) and the constant domains are derived from an Ig from a different species (often a human Ig). - A later European patent application, EP-A-0 125 023 (Genentech), relates to much the same subject as the Boss application, but also relates to the production by recombinant DNA technology of other variations of Ig-type molecules.
- EP-
A-0 194 276 (Neuberger) discloses not only chimeric antibodies of the type disclosed in the Boss application but also chimeric antibodies in which some or all of the constant domains have been replaced by non-Ig derived protein sequences. For instance, the heavy chain CH2 and CH3 domains may be replaced by protein sequences derived from an enzyme or a protein toxin. - EP-A-0 239 400 (Winter) discloses a different approach to the production of Ig molecules. In this approach, only the CDRs from a first type of Ig are grafted onto a second type of Ig in place of its normal CDRs. The Ig molecule thus produced is predominantly of the second type, since the CDRs form a relatively small part of the whole Ig. However, since the CDRs are the parts which define the specificity of the Ig, the Ig molecule thus produced has its specificity derived from the first Ig.
- Hereinafter, chimeric antibodies, CDR-grafted Igs, the altered antibodies described by Genentech, and fragments of such Igs such as F(ab′)2 and Fv fragments are referred to herein as modified antibodies.
- One of the main reasons for all the activity in the Ig field using recombinant DNA technology is the desire to use Igs in therapy. It is well known that, using the hybridoma technique developed by Kohler and Milstein, it is possible to produce monoclonal antibodies (MAbs) of almost any specificity. Thus, MAbs directed against cancer antigens have been produced. It is envisaged that these MAbs could be covalently attached or fused to toxins to provide “magic bullets” for use in cancer therapy. MAbs directed against normal tissue or cell surface antigens have also been produced. Labels can be attached to these so that they can be used for in vivo imaging.
- The major obstacle to the use of such MAbs in therapy or in vivo diagnosis is that the vast majority of MAbs which are produced are of rodent, in particular mouse, origin. It is very difficult to produce human MAbs. Since most MAbs are derived from non-human species, they are antigenic in humans. Thus, administration of these MAbs to humans generally results in an anti-Ig response being mounted by the human. Such a response can interfere with therapy or diagnosis, for instance by destroying or clearing the antibody quickly, or can cause allergic reactions or immune complex hypersensitivity which has adverse effects on the patient.
- The production of modified Igs has been proposed to ensure that the Ig administered to a patient is as “human” as possible, but still retains the appropriate specificity. It is therefore expected that modified Igs will be as effective as the MAb from which the specificity is derived but at the same time not very antigenic. Thus, it should be possible to use the modified Ig a reasonable number of times in a treatment or diagnosis regime.
- At the level of the gene, it is known that heavy chain variable domains are encoded by a “rearranged” gene which is built from three gene segments: an “unrearranged” VH gene (encoding the N-terminal three framework regions, first two complete CDRs and the first part of the third CDR), a diversity (DH)-segment (DH) (encoding the central portion of the third CDR) and a joining segment (JH) (encoding the last part of the third CDR and the fourth framework region). In the maturation of B-cells, the genes rearrange so that each unrearranged VH gene is linked to one DH gene and one JH gene. The rearranged gene corresponds to VH-DH-JH. This rearranged gene is linked to a gene which encodes the constant portion of the Ig chain.
- For light chains, the situation is similar, except that for light chains there is no diversity region. Thus light chain variable domains are encoded by an “unrearranged” VL gene and a JL gene. There are two types of light chains, kappa (κ) or lambda (λ), which are built respectively from unrearranged Vκ genes and Jκ segments, and from unrearranged Vλ genes and Jλ segments.
- Previous work has shown that it is necessary to have two variable domains in association together for efficient binding. For example, the associated heavy and light chain variable domains were shown to contain the antigen binding site [1]. This assumption is borne out by X-ray crystallographic studies of crystallized antibody/antigen complexes [2-6] which show that both the heavy and light chains of the antibody's variable domains contact the antigen. The expectation that association of heavy and light chain variable domains is necessary for efficient antigen binding underlies work to co-secrete these domains from bacteria [1], and to link the domains together by a short section of polypeptide as in the single chain antibodies [8, 9].
- Binding of isolated heavy and light chains had also been detected. However the evidence suggested strongly that this was a property of heavy or light chain dimmers. Early work, mainly with polyclonal antibodies, in which antibody heavy and light chains had been separated under denaturing conditions [10] suggested that isolated antibody heavy chains could bind to protein antigens [11] or hapten [12]. The binding of protein antigen was not characterized, but the hapten-binding affinity of the heavy chain fragments was reduced by two orders of magnitude [12] and the number of hapten molecules binding were variously estimated as 0.14 or 0.37 [13] or 0.26 [14] per isolated heavy chain. Furthermore binding of haptens was shown to be a property of dimeric heavy or dimeric light chains [14]. Indeed light chain dimers have been crystallized. It has been shown that in light chain dimers the two chains form a cavity which is able to bind to a single molecule of hapten [15].
- This confirms the assumption that, in order to obtain efficient binding, it is necessary to have a dimer, and preferably a heavy chain/light chain dimer, containing the respective variable domains. This assumption also underlies the teaching of the patent references cited above, wherein the intention is always to produce dimeric, and preferably heavy/light chain dimeric, molecules.
- It has now been discovered, contrary to expectations, that isolated Ig heavy chain variable domains can bind to antigen in a 1:1 ratio and with binding constants of equivalent magnitude to those of complete antibody molecules. In view of what was known up until now and in view of the assumptions made by those skilled in the art, this is highly surprising.
- Therefore, according to a first aspect of the present invention, there is provided a single domain ligand consisting of at least part of the variable domain of one chain of a molecule from the Ig superfamily.
- Preferably, the ligand consists of the variable domain of an Ig light, or, most preferably, heavy chain.
- The ligand may be produced by any known technique, for instance by controlled cleavage of Ig superfamily molecules or by peptide synthesis. However, preferably the ligand is produced by recombinant DNA technology. For instance, the gene encoding the rearranged gene for a heavy chain variable domain may be produced, for instance by cloning or gene synthesis, and placed into a suitable expression vector. The expression vector is then used to transform a compatible host cell which is then cultured to allow the ligand to be expressed and, preferably, secreted.
- If desired, the gene for the ligand can be mutated to improve the properties of the expressed domain, for example to increase the yields of expression or the solubility of the ligand, to enable the ligand to bind better, or to introduce a second site for covalent attachment (by introducing chemically reactive residues such as cysteine and histidine) or non-covalent binding of other molecules. In particular it would be desirable to introduce a second site for binding to serum components, to prolong the residence time of the domains in the serum; or for binding to molecules with effector functions, such as components of complement, or receptors on the surfaces of cells.
- Thus, hydrophobic residues which would normally be at the interface of the heavy chain variable domain with the light chain variable domain could be mutated to more hydrophilic residues to improve solubility; residues in the CDR loops could be mutated to improve antigen binding; residues on the other loops or parts of the β-sheet could be mutated to introduce new binding activities. Mutations could include single point mutations, multiple point mutations or more extensive changes and could be introduced by any of a variety of recombinant DNA methods, for example gene synthesis, site directed mutagenesis or the polymerase chain reaction.
- Since the ligands of the present invention have equivalent binding affinity to that of complete Ig molecules, the ligands can be used in many of the ways as are Ig molecules or fragments. For example, Ig molecules have been used in therapy (such as in treating cancer, bacterial and viral diseases), in diagnosis (such as pregnancy testing), in vaccination (such as in producing anti-idiotypic antibodies which mimic antigens), in modulation of activities of hormones or growth factors, in detection, in biosensors and in catalysis.
- It is envisaged that the small size of the ligands of the present invention may confer some advantages over complete antibodies, for example, in neutralizing the activity of low molecular weight drugs (such as dioxin) and allowing their filtration from the kidneys with drug attached; in penetrating tissues and tumors; in neutralizing viruses by binding to small conserved regions on the surfaces of viruses such as the “canyon” sites of viruses [16]; in high resolution epitope mapping of proteins; and in vaccination by ligands which mimic antigens.
- The present invention also provides receptors comprising a ligand according to the first aspect of the invention linked to one or more of an effector molecule, a label, a surface, or one or more other ligands having the same or different specificity.
- A receptor comprising a ligand linked to an effector molecule may be of use in therapy. The effector molecule may be a toxin, such as ricin or pseudomonas exotoxin, an enzyme which is able to activate a prodrug, a binding partner or a radio-isotope. The radio-isotope may be directly linked to the ligand or may be attached thereto by a chelating structure which is directly linked to the ligand. Such ligands with attached isotopes are much smaller than those based on Fv fragments, and could penetrate tissues and access tumors more readily.
- A receptor comprising a ligand linked to a label may be of use in diagnosis. The label may be a heavy metal atom or a radio-isotope, in which case the receptor can be used for in vivo imaging using X-ray or other scanning apparatus. The metal atom or radio-isotope may be attached to the ligand either directly or via a chelating structure directly linked to the ligand. For in vitro diagnostic testing, the label may be a heavy metal atom, a radio-isotope, an enzyme, a fluorescent or colored molecule or a protein or peptide tag which can be detected by an antibody, an antibody fragment or another protein. Such receptors would be used in any of the known diagnostic tests, such as ELISA or fluorescence-linked assays.
- A receptor comprising a ligand linked to a surface, such as a chromatography medium, could be used for purification of other molecules by affinity chromatography. Linking of ligands to cells, for example to the outer membrane proteins of E. coli or to hydrophobic tails which localize the ligands in the cell membranes, could allow a simple diagnostic test in which the bacteria or cells would agglutinate in the presence of molecules bearing multiple sites for binding the ligand(s).
- Receptors comprising at least two ligands can be used, for instance, in diagnostic tests. The first ligand will bind to a test antigen and the second ligand will bind to a reporter molecule, such as an enzyme, a fluorescent dye, a colored dye, a radio-isotope or a colored-, fluorescently- or radio-labelled protein.
- Alternatively, such receptors may be useful in increasing the binding to an antigen. The first ligand will bind to a first epitope of the antigen and the second ligand will bind to a second epitope. Such receptors may also be used for increasing the affinity and specificity of binding to different antigens in close proximity on the surface of cells. The first ligand will bind to the first antigen and the second epitope to the second antigen: strong binding will depend on the co-expression of the epitopes on the surface of the cell. This may be useful in therapy of tumors, which can have elevated expression of several surface markers. Further ligands could be added to further improve binding or specificity. Moreover, the use of strings of ligands, with the same or multiple specificities, creates a larger molecule which is less readily filtered from the circulation by the kidney.
- For vaccination with ligands which mimic antigens, the use of strings of ligands may prove more effective than single ligands, due to repetition of the immunizing epitopes.
- If desired, such receptors with multiple ligands could include effector molecules or labels so that they can be used in therapy or diagnosis as described above.
- The ligand may be linked to the other part of the receptor by any suitable means, for instance by covalent or non-covalent chemical linkages. However, where the receptor comprises a ligand and another protein molecule, it is preferred that they are produced by recombinant DNA technology as a fusion product. If necessary, a linker peptide sequence can be placed between the ligand and the other protein molecule to provide flexibility.
- The basic techniques for manipulating Ig molecules by recombinant DNA technology are described in the patent references cited above. These may be adapted in order to allow for the production of ligands and receptors according to the invention by means of recombinant DNA technology.
- Preferably, where the ligand is to be used for in vivo diagnosis or therapy in humans, it is humanized, for instance by CDR replacement as described in EP-A-0 239 400.
- In order to obtain a DNA sequence encoding a ligand, it is generally necessary firstly to produce a hybridoma which secretes an appropriate MAb. This can be a very time consuming method. Once an immunized animal has been produced, it is necessary to fuse separated spleen cells with a suitable myeloma cell line, grow up the cell lines thus produced, select appropriate lines, reclone the selected lines and reselect. This can take some long time. This problem also applies to the production of modified Igs.
- A further problem with the production of ligands, and also receptors according to the invention and modified Igs, by recombinant DNA technology is the cloning of the variable domain encoding sequences from the hybridoma which produces the MAb from which the specificity is to be derived. This can be a relatively long method involving the production of a suitable probe, construction of a clone library from cDNA or genomic DNA, extensive probing of the clone library, and manipulation of any isolated clones to enable the cloning into a suitable expression vector. Due to the inherent variability of the DNA sequences encoding Ig variable domains, it has not previously been possible to avoid such time consuming work. It is therefore a further aim of the present invention to provide a method which enables substantially any sequence encoding an Ig superfamily molecule variable domain (ligand) to be cloned in a reasonable period of time.
- According to another aspect of the present invention therefore, there is provided a method of cloning a sequence (the target sequence) which encodes at least part of the variable domain of an Ig superfamily molecule, which method comprises:
- (a) providing a sample of double stranded (ds) nucleic acid which contains the target sequence;
- (b) denaturing the sample so as to separate the two strands;
- (c) annealing to the sample a forward and a back oligonucleotide primer, the forward primer being specific for a sequence at or adjacent the 3′ end of the sense strand of the target sequence, the back primer being specific for a sequence at or adjacent the 3′ end of the antisense strand of the target sequence, under conditions which allow the primers to hybridize to the nucleic acid at or adjacent the target sequence;
- (d) treating the annealed sample with a DNA polymerase enzyme in the presence of deoxynucleoside triphosphates under conditions which cause primer extension to take place; and
- (e) denaturing the sample under conditions such that the extended primers become separated from the target sequence.
- Preferably, the method of the present invention further includes the step (f) of repeating steps (c) to (e) on the denatured mixture a plurality of times.
- Preferably, the method of the present invention is used to clone complete variable domains from Ig molecules, most preferably from Ig heavy chains. In the most preferred instance, the method will produce a DNA sequence encoding a ligand according to the present invention.
- In step (c) recited above, the forward primer becomes annealed to the sense strand of the target sequence at or adjacent the 3′ end of the strand. In a similar manner, the back primer becomes annealed to the antisense strand of the target sequence at or adjacent the 3′ end of the strand. Thus, the forward primer anneals at or adjacent the region of the ds nucleic acid which encodes the C-terminal end of the variable region or domain. Similarly, the back primer anneals at or adjacent the region of the ds nucleic acid which encodes the N-terminal end of the variable domain.
- In step (d), nucleotides are added onto the 3′ end of the forward and back primers in accordance with the sequence of the strand to which they are annealed. Primer extension will continue in this manner until stopped by the beginning of the denaturing step (e). It must therefore be ensured that step (d) is carried out for a long enough time to ensure that the primers are extended so that the extended strands totally overlap one another.
- In step (e), the extended primers are separated from the ds nucleic acid. The ds nucleic acid can then serve again as a substrate to which further primers can anneal. Moreover, the extended primers themselves have the necessary complementary sequences to enable the primers to anneal thereto.
- During further cycles, if step (f) is used, the amount of extended primers will increase exponentially so that at the end of the cycles there will be a large quantity of cDNA having sequences complementary to the sense and antisense strands of the target sequence. Thus, the method of the present invention will result in the accumulation of a large quantity of cDNA which can form ds cDNA encoding at least part of the variable domain.
- As will be apparent to the skilled person, some of the steps in the method may be carried out simultaneously or sequentially as desired.
- The forward and back primers may be provided as isolated oligonucleotides, in which case only two oligonucleotides will be used. However, alternatively the forward and back primers may each be supplied as a mixture of closely related oligonucleotides. For instance, it may be found that at a particular point in the sequence to which the primer is to anneal, there is the possibility of nucleotide variation. In this case a primer may be used for each possible nucleotide variation. Furthermore it may be possible to use two or more sets of “nested” primers in the method to enhance the specific cloning of variable region genes.
- The method described above is similar to the method described by Saiki et al. [17]. A similar method is also used in the methods described in EP-
A-0 200 362. In both cases the method described is carried out using primers which are known to anneal efficiently to the specified nucleotide sequence. In neither of these disclosures was it suggested that the method could be used to clone Ig parts of variable domain encoding sequences, where the target sequence contains inherently highly variable areas. - The ds nucleic acid sequence used in the method of the present invention may be derived from mRNA. For instance, RNA may be isolated in known manner from a cell or cell line which is known to produce Igs. mRNA may be separated from other RNA by oligo-dT chromatography. A complementary strand of cDNA may then be synthesized on the mRNA template, using reverse transcriptase and a suitable primer, to yield an RNA/DNA heteroduplex. A second strand of DNA can be made in one of several ways, for example, by priming with RNA fragments of the mRNA strand (made by incubating RNA/DNA heteroduplex with RNase H) and using DNA polymerase, or by priming with a synthetic oligodeoxynucleotide primer which anneals to the 3′ end of the first strand and using DNA polymerase. It has been found that the method of the present invention can be carried out using ds cDNA prepared in this way.
- When making such ds cDNA, it is possible to use a forward primer which anneals to a sequence in the CH1 domain (for a heavy chain variable domain) or the Cλ or Cκ domain (for a light chain variable domain). These will be located in close enough proximity to the target sequence to allow the sequence to be cloned.
- The back primer may be one which anneals to a sequence at the N-terminal end of the VH1, Vκ or V λ domain. The back primer may consist of a plurality of primers having a variety of sequences designed to be complementary to the various families of VH1, Vκ or Vλ sequences known. Alternatively the back primer may be a single primer having a consensus sequence derived from all the families of variable region genes.
- Surprisingly, it has been found that the method of the present invention can be carried out using genomic DNA. If genomic DNA is used, there is a very large amount of DNA present, including actual coding sequences, introns and untranslated sequences between genes. Thus, there is considerable scope for non-specific annealing under the conditions used. However, it has surprisingly been found that there is very little non-specific annealing. It is therefore unexpected that it has proved possible to clone the genes of Ig-variable domains from genomic DNA.
- Under some circumstances the use of genomic DNA may prove advantageous compared with use of mRNA, as the mRNA is readily degraded, and especially difficult to prepare from clinical samples of human tissue.
- Thus, in accordance with an aspect of the present invention, the ds nucleic acid used in step (a) is genomic DNA.
- When using genomic DNA as the ds nucleic acid source, it will not be possible to use as the forward primer an oligonucleotide having a sequence complementary to a sequence in a constant domain. This is because, in genomic DNA, the constant domain genes are generally separated from the variable domain genes by a considerable number of base pairs. Thus, the site of annealing would be too remote from the sequence to be cloned.
- It should be noted that the method of the present invention can be used to clone both rearranged and unrearranged variable domain sequences from genomic DNA. It is known that in germ line genomic DNA the three genes, encoding the VH, DH and JH respectively, are separated from one another by considerable numbers of base pairs. On maturation of the immune response, these genes are rearranged so that the VH, DH and JH genes are fused together to provide the gene encoding the whole variable domain (see
FIG. 1 ). By using a forward primer specific for a sequence at or adjacent the 3′ end of the sense strand of the genomic “unrearranged” VH gene, it is possible to clone the “unrearranged” VH gene alone, without also cloning the DH and JH genes. This can be of use in that it will then be possible to fuse the VH gene onto pre-cloned or synthetic DH and DH genes. In this way, rearrangement of the variable domain genes can be carried out in vitro. - The oligonucleotide primers used in step (c) may be specifically designed for use with a particular target sequence. In this case, it will be necessary to sequence at least the 5′ and 3′ ends of the target sequence so that the appropriate oligonucleotides can be synthesized. However, the present inventors have discovered that it is not necessary to use such specifically designed primers. Instead, it is possible to use a species specific general primer or a mixture of such primers for annealing to each end of the target sequence. This is not particularly surprising as regards the 3′ end of the target sequence. It is known that this end of the variable domain encoding sequence leads into a segment encoding JH which is known to be relatively conserved. However, it was surprisingly discovered that, within a single species, the sequence at the 5′ end of the target sequence is sufficiently well conserved to enable a species specific general primer or a mixture thereof to be designed for the 5′ end of the target sequence.
- Therefore according to a preferred aspect of the present invention, in step (c) the two primers which are used are species specific general primers, whether used as single primers or as mixtures of primers. This greatly facilitates the cloning of any undetermined target sequence since it will avoid the need to carry out any sequencing on the target sequence in order to produce target sequence-specific primers. Thus the method of this aspect of the invention provides a general method for cloning variable region or domain encoding sequences of a particular species.
- Once the variable domain gene has been cloned using the method described above, it may be directly inserted into an expression vector, for instance using the PCR reaction to paste the gene into a vector.
- Advantageously, however, each primer includes a sequence including a restriction enzyme recognition site. The sequence recognized by the restriction enzyme need not be in the part of the primer which anneals to the ds nucleic acid, but may be provided as an extension which does not anneal. The use of primers with restriction sites has the advantage that the DNA can be cut with at least one restriction enzyme which leaves 3′ or 5′ overhanging nucleotides. Such DNA is more readily cloned into the corresponding sites on the vectors than blunt end fragments taken directly from the method. The ds cDNA produced at the end of the cycles will thus be readily insertable into a cloning vector by use of the appropriate restriction enzymes. Preferably the choice of restriction sites is such that the ds cDNA is cloned directly into an expression vector, such that the ligand encoded by the gene is expressed. In this case the restriction site is preferably located in the sequence which is annealed to the ds nucleic acid.
- Since the primers may not have a sequence exactly complementary to the target sequence to which it is to be annealed, for instance because of nucleotide variations or because of the introduction of a restriction enzyme recognition site, it may be necessary to adjust the conditions in the annealing mixture to enable the primers to anneal to the ds nucleic acid. This is well within the competence of the person skilled in the art and needs no further explanation.
- In step (d), any DNA polymerase may be used. Such polymerases are known in the art and are available commercially. The conditions to be used with each polymerase are well known and require no further explanation here. The polymerase reaction will need to be carried out in the presence of the four nucleoside triphosphates. These and the polymerase enzyme may already be present in the sample or may be provided afresh for each cycle.
- The denaturing step (e) may be carried out, for instance, by heating the sample, by use of chaotropic agents, such as urea or guanidine, or by the use of changes in ionic strength or pH. Preferably, denaturing is carried out by heating since this is readily reversible. Where heating is used to carry out the denaturing, it will be usual to use a thermostable DNA polymerase, such as Taq polymerase, since this will not need replenishing at each cycle.
- If heating is used to control the method, a suitable cycle of heating comprises denaturation at about 95° C. for about 1 minute, annealing at from 30° C. to 65° C. for about 1 minute and primer extension at about 75° C. for about 2 minutes. To ensure that elongation and renaturation is complete, the mixture after the final cycle is preferably held at about 60° C. for about 5 minutes.
- The product ds cDNA may be separated from the mixture for instance by gel electrophoresis using agarose gels. However, if desired, the ds cDNA may be used in unpurified form and inserted directly into a suitable cloning or expression vector by conventional methods. This will be particularly easy to accomplish if the primers include restriction enzyme recognition sequences.
- The method of the present invention may be used to make variations in the sequences encoding the variable domains. For example this may be achieved by using a mixture of related oligonucleotide primers as at least one of the primers. Preferably the primers are particularly variable in the middle of the primer and relatively conserved at the 5′ and 3′ ends. Preferably the ends of the primers are complementary to the framework regions of the variable domain, and the variable region in the middle of the primer covers all or part of a CDR. Preferably a forward primer is used in the area which forms the third CDR. If the method is carried out using such a mixture of oligonucleotides, the product will be a mixture of variable domain encoding sequences. Moreover, variations in the sequence may be introduced by incorporating some mutagenic nucleotide triphosphates in step (d), such that point mutations are scattered throughout the target region. Alternatively such point mutations are introduced by performing a large number of cycles of amplification, as errors due to the natural error rate of the DNA polymerase are amplified, particularly when using high concentrations of nucleoside triphosphates.
- The method of this aspect of the present invention has the advantage that it greatly facilitates the cloning of variable domain encoding sequences directly from mRNA or genomic DNA. This in turn will facilitate the production of modified Ig-type molecules by any of the prior art methods referred to above. Further, target genes can be cloned from tissue samples containing antibody producing cells, and the genes can be sequenced. By doing this, it will be possible to look directly at the immune repertoire of a patient. This “fingerprinting” of a patient's immune repertoire could be of use in diagnosis, for instance of auto-immune diseases.
- In the method for amplifying the amount of a gene encoding a variable domain, a single set of primers is used in several cycles of copying via the polymerase chain reaction. As a less preferred alternative, there is provided a second method which comprises steps (a) to (d) as above, which further includes the steps of:
- (g) treating the sample of ds cDNA with traces of DNAse in the presence of DNA polymerase I to allow nick translation of the DNA; and
- (h) cloning the ds cDNA into a vector.
- If desired, the second method may further include the steps of:
- (i) digesting the DNA of recombinant plasmids to release DNA fragments containing genes encoding variable domains; and
- (j) treating the fragments in a further set of steps (c) to (h).
- Preferably the fragments are separated from the vector and from other fragments of the incorrect size by gel electrophoresis.
- The steps (a) to (d) then (g) to (h) can be followed once, but preferably the entire cycle (c) to (d) and (g) to (h) is repeated at least once. In this way a priming step, in which the genes are specifically copied, is followed by a cloning step, in which the amount of genes is increased.
- In step (a) the ds cDNA is derived from mRNA. For Ig derived variable domains, the mRNA is preferably be isolated from lymphocytes which have been stimulated to enhance production of mRNA.
- In each step (c) the set of primers are preferably different from the previous step (c), so as to enhance the specificity of copying. Thus the sets of primers form a nested set. For example, for cloning of Ig heavy chain variable domains, the first set of primers may be located within the signal sequence and constant region, as described by Larrick et al., [18], and the second set of primers entirely within the variable region, as described by Orlandi et al., [19]. Preferably the primers of step (c) include restriction sites to facilitate subsequent cloning. In the last cycle the set of primers used in step (c) should preferably include restriction sites for introduction into expression vectors. In step (g) possible mismatches between the primers and the template strands are corrected by “nick translation”. In step (h), the ds cDNA is preferably cleaved with restriction enzymes at sites introduced into the primers to facilitate the cloning.
- According to another aspect of the present invention the product ds cDNA is cloned directly into an expression vector. The host may be prokaryotic or eukaryotic, but is preferably bacterial. Preferably the choice of restriction sites in the primers and in the vector, and other features of the vector will allow the expression of complete ligands, while preserving all those features of the amino acid sequence which are typical of the (methoded) ligands. For example, for expression of the rearranged variable genes, the primers would be chosen to allow the cloning of target sequences including at least all the three CDR sequences. The cloning vector would then encode a signal sequence (for secretion of the ligand), and sequences encoding the N-terminal end of the first framework region, restriction sites for cloning and then the C-terminal end of the last (fourth) framework region.
- For expression of unrearranged VH genes as part of complete ligands, the primers would be chosen to allow the cloning of target sequences including at least the first two CDRs. The cloning vector could then encode signal sequence, the N-terminal end of the first framework region, restriction sites for cloning and then the C-terminal end of the third framework region, the third CDR and fourth framework region.
- Primers and cloning vectors may likewise be devised for expression of single CDRs, particularly the third CDR, as parts of complete ligands. The advantage of cloning repertoires of single CDRs would permit the design of a “universal” set of framework regions, incorporating desirable properties such as solubility.
- Single ligands could be expressed alone or in combination with a complementary variable domain. For example, a heavy chain variable domain can be expressed either as an individual domain or, if it is expressed with a complementary light chain variable domain, as an antigen binding site. Preferably the two partners would be expressed in the same cell, or secreted from the same cell, and the proteins allowed to associate non-covalently to form an Fv fragment. Thus the two genes encoding the complementary partners can be placed in tandem and expressed from a single vector, the vector including two sets of restriction sites. Preferably the genes are introduced sequentially: for example the heavy chain variable domain can be cloned first and then the light chain variable domain. Alternatively the two genes are introduced into the vector in a single step, for example by using the polymerase chain reaction to paste together each gene with any necessary intervening sequence, as essentially described by Yon and Fried [29]. The two partners could be also expressed as a linked protein to produce a single chain Fv fragment, using similar vectors to those described above. As a further alternative the two genes may be placed in two different vectors, for example in which one vector is a phage vector and the other is a plasmid vector.
- Moreover, the cloned ds cDNA may be inserted into an expression vector already containing sequences encoding one or more constant domains to allow the vector to express Ig-type chains. The expression of Fab fragments, for example, would have the advantage over Fv fragments that the heavy and light chains would tend to associate through the constant domains in addition to the variable domains. The final expression product may be any of the modified Ig-type molecules referred to above.
- The cloned sequence may also be inserted into an expression vector so that it can be expressed as a fusion protein. The variable domain encoding sequence may be linked directly or via a linker sequence to a DNA sequence encoding any protein effector molecule, such as a toxin, enzyme, label or another ligand. The variable domain sequences may also be linked to proteins on the outer side of bacteria or phage. Thus, the method of this aspect of the invention may be used to produce receptors according to the invention.
- According to another aspect of the invention, the cloning of ds cDNA directly for expression permits the rapid construction of expression libraries which can be screened for binding activities. For Ig heavy and light chain variable genes, the ds cDNA may comprise variable genes isolated as complete rearranged genes from the animal, or variable genes built from several different sources, for example a repertoire of unrearranged VH genes combined with a synthetic repertoire of DH and JH genes. Preferably repertoires of genes encoding Ig heavy chain variable domains are prepared from lymphocytes of animals immunized with an antigen.
- The screening method may take a range of formats well known in the art. For example Ig heavy chain variable domains secreted from bacteria may be screened by binding to antigen on a solid phase, and detecting the captured domains by antibodies. Thus the domains may be screened by growing the bacteria in liquid culture and binding to antigen coated on the surface of ELISA plates. However, preferably bacterial colonies (or phage plaques) which secrete ligands (or modified ligands, or ligand fusions with proteins) are screened for antigen binding on membranes. Either the ligands are bound directly to the membranes (and for example detected with labelled antigen), or captured on antigen coated membranes (and detected with reagents specific for ligands). The use of membranes offers great convenience in screening many clones, and such techniques are well known in the art.
- The screening method may also be greatly facilitated by making protein fusions with the ligands, for example by introducing a peptide tag which is recognized by an antibody at the N-terminal or C-terminal end of the ligand, or joining the ligand to an enzyme which catalyses the conversion of a colorless substrate to a colored product. In the latter case, the binding of antigen may be detected simply by adding substrate. Alternatively, for ligands expressed and folded correctly inside eukaryotic cells, joining of the ligand and a domain of a transcriptional activator such as the GAL4 protein of yeast, and joining of antigen to the other domain of the GAL4 protein, could form the basis for screening binding activities, as described by Fields and Song [21].
- The preparation of proteins, or even cells with multiple copies of the ligands, may improve the avidity of the ligand for immobilized antigen, and hence the sensitivity of the screening method. For example, the ligand may be joined to a protein subunit of a multimeric protein, to a phage coat protein or to an outer membrane protein of E. coli such as ompA or lamB. Such fusions to phage or bacterial proteins also offers possibilities of selecting bacteria displaying ligands with antigen binding activities. For example such bacteria may be precipitated with antigen bound to a solid support, or may be subjected to affinity chromatography, or may be bound to larger cells or particles which have been coated with antigen and sorted using a fluorescence activated cell sorter (FACS). The proteins or peptides fused to the ligands are preferably encoded by the vector, such that cloning of the ds cDNA repertoire creates the fusion product.
- In addition to screening for binding activities of single ligands, it may be necessary to screen for binding or catalytic activities of associated ligands, for example, the associated Ig heavy and light chain variable domains. For example, repertoires of heavy and light chain variable genes may be cloned such that two domains are expressed together. Only some of the pairs of domains may associate, and only some of these associated pairs may bind to antigen. The repertoires of heavy and light chain variable domains could be cloned such that each domain is paired at random. This approach may be most suitable for isolation of associated domains in which the presence of both partners is required to form a cleft. Alternatively, to allow the binding of hapten. Alternatively, since the repertoires of light chain sequences are less diverse than those of heavy chains, a small repertoire of light chain variable domains, for example including representative members of each family of domains, may be combined with a large repertoire of heavy chain variable domains.
- Preferably however, a repertoire of heavy chain variable domains is screened first for antigen binding in the absence of the light chain partner, and then only those heavy chain variable domains binding to antigen are combined with the repertoire of light chain variable domains. Binding of associated heavy and light chain variable domains may be distinguished readily from binding of single domains, for example by fusing each domain to a different C-terminal peptide tag which are specifically recognized by different monoclonal antibodies.
- The hierarchical approach of first cloning heavy chain variable domains with binding activities, then cloning matching light chain variable domains may be particularly appropriate for the construction of catalytic antibodies, as the heavy chain may be screened first for substrate binding. A light chain variable domain would then be identified which is capable of association with the heavy chain, and “catalytic” residues such as cysteine or histidine (or prosthetic groups) would be introduced into the CDRs to stabilize the transition state or attack the substrate, as described by Baldwin and Schultz [22].
- Although the binding activities of non-covalently associated heavy and light chain variable domains (Fv fragments) may be screened, suitable fusion proteins may drive the association of the variable domain partners. Thus Fab fragments are more likely to be associated than the Fv fragments, as the heavy chain variable domain is attached to a single heavy chain constant domain, and the light chain variable domain is attached to a single light chain variable domain, and the two constant domains associate together.
- Alternatively the heavy and light chain variable domains are covalently linked together with a peptide, as in the single chain antibodies, or peptide sequences attached, preferably at the C-terminal end which will associate through forming cysteine bonds or through non-covalent interactions, such as the introduction of “leucine zipper” motifs. However, in order to isolate pairs of tightly associated variable domains, the Fv fragments are preferably used.
- The construction of Fv fragments isolated from a repertoire of variable region genes offers a way of building complete antibodies, and an alternative to hybridoma technology. For example by attaching the variable domains to light or suitable heavy chain constant domains, as appropriate, and expressing the assembled genes in mammalian cells, complete antibodies may be made and should possess natural effector functions, such as complement lysis. This route is particularly attractive for the construction of human monoclonal antibodies, as hybridoma technology has proved difficult, and for example, although human peripheral blood lymphocytes can be immortalized with Epstein Barr virus, such hybridomas tend to secrete low affinity IgM antibodies.
- Moreover, it is known that immunological mechanisms ensure that lymphocytes do not generally secrete antibodies directed against host-proteins. However it is desirable to make human antibodies directed against human proteins, for example to human cell surface markers to treat cancers, or to histocompatibility antigens to treat auto-immune diseases. The construction of human antibodies built from the combinatorial repertoire of heavy and light chain variable domains may overcome this problem, as it will allow human antibodies to be built with specificities which would normally have been eliminated.
- The method also offers a new way of making bispecific antibodies. Antibodies with dual specificity can be made by fusing two hybridomas of different specificities, so as to make a hybrid antibody with an Fab arm of one specificity, and the other Fab arm of a second specificity. However the yields of the bispecific antibody are low, as heavy and light chains also find the wrong partners. The construction of Fv fragments which are tightly associated should preferentially drive the association of the correct pairs of heavy with light chains. (It would not assist in the correct pairing of the two heavy chains with each other.) The improved production of bispecific antibodies would have a variety of applications in diagnosis and therapy, as is well known.
- Thus the invention provides a species specific general oligonucleotide primer or a mixture of such primers useful for cloning variable domain encoding sequences from animals of that species. The method allows a single pair or pair of mixtures of species specific general primers to be used to clone any desired antibody specificity from that species. This eliminates the need to carry out any sequencing of the target sequence to be cloned and the need to design specific primers for each specificity to be recovered.
- Furthermore it provides for the construction of repertoires of variable genes, for the expression of the variable genes directly on cloning, for the screening of the encoded domains for binding activities and for the assembly of the domains with other variable domains derived from the repertoire.
- Thus the use of the method of the present invention will allow for the production of heavy chain variable domains with binding activities and variants of these domains. It allows for the production of monoclonal antibodies and bispecific antibodies, and will provide an alternative to hybridoma technology. For instance, mouse splenic ds mRNA or genomic DNA may be obtained from a hyper-immunized mouse. This could be cloned using the method of the present invention and then the cloned ds DNA inserted into a suitable expression vector. The expression vector would be used to transform a host cell, for instance a bacterial cell, to enable it to produce an Fv fragment or a Fab fragment. The Fv or Fab fragment would then be built into a monoclonal antibody by attaching constant domains and expressing it in mammalian cells.
- In the Examples described below, the following oligonucleotide primers, or mixed primers were used. Their locations are marked on
FIG. 1 and sequences are as follows: -
VH1FOR 5′ TGAGGAGACGGTGACCGTGGTCCCTTGGCCCCAG 3′; VH1FOR-2 5′ TGAGGAGACGGTGACCGTGGTCCCTTGGCCCC 3′; Hu1VHFOR 5′ CTTGGTGGAGGCTGAGGAGACGGTGACC 3′; Hu2VHFOR 5′ CTTGGTGGAGGCTGAGGAGACGGTGACC 3′; Hu3VHFOR 5′ CTTGGTGGATGCTGAGGAGACGGTGACC 3′; Hu4VHFOR 5′ CTTGGTGGATGCTGATGAGACGGTGACC 3′; MOJH1FOR 5′ TGAGGAGACGGTGACCGTGGTCCCTGCGCCCCAG 3′; MOJH2FOR 5′ TGAGGAGACGGTGACCGTGGTGCCTTGGCCCCAG 3′; MOJH3FOR 5′ TGCAGAGACGGTGACCAGAGTCCCTTGGCCCCAG 3′; MOJH4FOR 5′ TGAGGAGACGGTGACCGAGGTTCCTTGACCCCAG 3′; HUJH1FOR 5′ TGAGGAGACGGTGACCAGGGTGCCCTGGCCCCAG 3′; HUJH2FOR 5′ TGAGGAGACGGTGACCAGGGTGCCACGGCCCCAG 3′; HUJH4FOR 5′ TGAGGAGACGGTGACCAGGGTTCCTTGGCCCCAG 3′; VK1FOR 5′ GTTAGATCTCCAGCTTGGTCCC 3′; VK2FOR 5′ CGTTAGATCTCCAGCTTGGTCCC 3′; VK3FOR 5′ CCGTTTCAGCTCGAGCTTGGTCCC 3′; MOJK1FOR 5′ CGTTAGATCTCCAGCTTGGTGCC 3′; MOJK3FOR 5′ GGTTAGATCTCCAGTCTGGTCCC 3′; MOJK4FOR 5′ CGTTAGATCTCCAACTTTGTCCC 3′; HUJK1FOR 5′ CGTTAGATCTCCACCTTGGTCCC 3′; HUJK3FOR 5′ CGTTAGATCTCCACTTTGGTCCC 3′; HUJK4FOR 5′ CGTTAGATCTCCACCTTGGTCCC 3′; HUJK5FOR 5′ CGTTAGATCTCCAGTCGTGTCCC 3′; VH1BACK 5′ AGGT(C/G)(C/A)A(G/A)CTGCAG(G/C)AGTC(T/A)GG 3′; Hu2VHIBACK: 5′ CAGGTGCAGCTGCAGCAGTCTGG 3′; HuVHIIBACK: 5′ CAGGTGCAGCTGCAGGAGTCGGG 3′; Hu2VHIIIBACK: 5′ GAGGTGCAGCTGCAGGAGTCTGG 3′; HuVHIVBACK: 5′ CAGGTGCAGCTGCAGCAGTCTGG 3′; MOVHIBACK 5′ AGGTGCAGCTGCAGGAGTCAG 3′; MOVHIIABACK 5′ AGGTCCAGCTGCAGCA(G/A)TCTGG 3′; MOVHIIBBACK 5′ AGGTCCAACTGCAGCAGCCTGG 3′; MOVHIIBACK 5′ AGGTGAAGCTGCAGGAGTCTGG 3′; VK1BACK 5′ GACATTCAGCTGACCCAGTCTCCA 3′; VK2BACK 5′ GACATTGAGCTCACCCAGTCTCCA 3′; MOVKIIABACK 5′ GATGTTCAGCTGACCCAAACTCCA 3′ MOVKIIBBACK 5′ GATATTCAGCTGACCCAGGATGAA 3′; HuHep1FOR 5′ C(A/G)(C/G)TGAGCTCACTGTGTCTCTCGCACA 3′; HuOcta1BACK 5′ CGTGAATATGCAAATAA 3′; HUOcta2BACK 5′ AGTAGGAGACATGCAAAT 3′; and HuOcta3BACK 5′ CACCACCCACATGCAAAT 3′; VHMUT1 5′ GGAGACGGTGACCGTGGTCCCTTGGCCCCAGTAGTCAAGNNNNNNNN NNNNCTCTCTGGC 3′ (where N is an equimolar mixture of T, C, G and A) M13 pRIMER 5′ AACAGCTATGACCATG 3′ (New England Biolabs *1201) - VH1FOR is designed to anneal with the 3′ end of the sense strand of any mouse heavy chain variable domain encoding sequence. It contains a BstEII recognition site. VK1FOR is designed to anneal with the 3′ end of the sense strand of any mouse kappa-type light chain variable domain encoding sequence and contains a BglII recognition site. VH1BACK is designed to anneal with the 3′ end of the antisense strand of any mouse heavy chain variable domain and contains a PstI recognition site. VK1BACK is designed to anneal with the 3′ end of the antisense strand of any mouse kappa-type light chain variable domain encoding sequence and contains a PvuII recognition site.
- In this Example five mouse hybridomas were used as a source of ds nucleic acid. The hybridomas produce monoclonal antibodies (MAbs) designated MBr1 [23], BW431/26 [24], BW494/32 [25], BW250/183 [24,26] and BW704/152 [27]. MAb MBr1 is particularly interesting in that it is known to be specific for a saccharide epitope on a human mammary carcinoma line MCF-7 [28].
- Cloning Via mRNA
- Each of the five hybridomas referred to above was grown up in roller bottles and about 5×108 cells of each hybridoma were used to isolate RNA. mRNA was separated from the isolated RNA using oligodT cellulose [29]. First strand cDNA was synthesized according to the procedure described by Maniatis et al. [30] as set out below.
- In order to clone the heavy chain variable domain encoding sequence, a 50 μl reaction solution which contains 10 μg mRNA, 20 pmole VH1FOR primer, 250 μM each of dATP, dTTP, dCTP and dGTP, 10 mM dithiothreitol (DTT), 100 mM Tris.HCl, 10 MM MgCl2 and 140 mM KCl, adjusted to pH 8.3 was prepared. The reaction solution was heated at 70° C. for ten minutes and allowed to cool to anneal the primer to the 3′ end of the variable domain encoding sequence in the mRNA. To the reaction solution was then added 46 units of reverse transcriptase (Anglian Biotec) and the solution was then incubated at 42° C. for 1 hour to cause first strand cDNA synthesis.
- In order to clone the light chain variable domain encoding sequence, the same procedure as set out above was used except that the VK1FOR primer was used in place of the VH1 FOR primer.
- Amplification from RNA/DNA Hybrid
- Once the ds RNA/DNA hybrids had been produced, the variable domain encoding sequences were amplified as follows. For heavy chain variable domain encoding sequence amplification, a 50 μl reaction solution containing 5 μl of the ds RNA/DNA hybrid-containing solution, 25 pmole each of VH1FOR and VH1BACK primers, 250 μM of dATP, dTTP, dCTP and dGTP, 67 mM Tris.HCl, 17 mM ammonium sulphate, 10 mM MgCl2, 200 μg/ml gelatine and 2 units Taq polymerase (Cetus) was prepared. The reaction solution was overlaid with paraffin oil and subjected to 25 rounds of temperature cycling using a Techne PHC-1 programmable heating block. Each cycle consisted of 1 minute and 95° C. (to denature the nucleic acids), 1 minute at 30° C. (to anneal the primers to the nucleic acids) and 2 minutes at 72° C. (to cause elongation from the primers). After the 25 cycles, the reaction solution and the oil were extracted twice with ether, once with phenol and once with phenol/CHCl3. Thereafter ds cDNA was precipitated with ethanol. The precipitated ds cDNA was then taken up in 50 μl of water and frozen.
- The procedure for light chain amplification was exactly as described above, except that the VK1FOR and VK1BACK primers were used in place of the VH1FOR and VH1BACK primers respectively.
- 5 μl of each sample of amplified cDNA was fractionated on 2% agarose gels by electrophoresis and stained with ethidium bromide. This showed that the amplified ds cDNA gave a major band of the expected size (about 330 bp). (However the band for VK 0 DNA of MBr1 was very weak. It was therefore excised from the gel and reamplified in a second round.) Thus by this simple procedure, reasonable quantities of ds DNA encoding the light and heavy chain variable domains of the five MAbs were produced.
- Heavy Chain Vector Construction
- A BstEII recognition site was introduced into the vector M13-HuVHNP [31] by site directed mutagenesis [32,33] to produce the vector M13-VHPCR1 (
FIGS. 2 and 3 ). - Each amplified heavy chain variable domain encoding sequence was digested with the restriction enzymes PstI and BstEII. The fragments were phenol extracted, purified on 2% low melting point agarose gels and force cloned into vector M13-VHPCR1 which had been digested with PstI and BstEII and purified on an 0.8% agarose gel. Clones containing the variable domain inserts were identified directly by sequencing [34] using primers based in the 3′ non-coding variable gene in the M13-VHPCR1 vector.
- There is an internal PstI site in the heavy chain variable domain encoding sequences of BW431/26. This variable domain encoding sequence was therefore assembled in two steps. The 3′ PstI-BstEII fragment was first cloned into M13-VHPCR1, followed in a second step by the 5′ PstI fragment.
- Light Chain Vector Construction
- Vector M13 mp 18 [35] was cut with PvuII and the vector backbone was blunt ligated to a synthetic HindIII-BamHI polylinker. Vector M13-HuVKLYS [36] was digested with HindIII and BamHI to isolate the HuVKLYS gene. This HindIII-BamHI fragment was then inserted into the HindIII-BamHI polylinker site to form a vector M13-VKPCR1 which lacks any PvuII sites in the vector backbone (
FIGS. 4 and 5 ). This vector was prepared in E. coli JM110 [22] to avoid dam methylation at the BclI site. - Each amplified light chain variable domain encoding sequence was digested with PvuII and BglII. The fragments were phenol extracted, purified on 2% low melting point agarose gels and force cloned into vector M13-VKPCR1 which had been digested with PvuII and BclI, purified on an 0.8% agarose gel and treated with calf intestinal phosphatase. Clones containing the light chain variable region inserts were identified directly by sequencing [34] using primers based in the 3′ non-coding region of the variable domain in the M13-VKPCR1 vector.
- The nucleotide sequences of the MBr1 heavy and light chain variable domains are shown in
FIG. 6 with part of the flanking regions of the M13-VHPCR1 and M13-VKPCR1 vectors. - Antibody Expression
- The HindIII-BamHI fragment carrying the MBr1 heavy chain variable domain encoding sequence in M13-VHPCR1 was recloned into a pSV-gpt vector with human γ1 constant regions [37] (
FIG. 7 ). The MBr1 light chain variable domain encoding sequence in M13-VKPCR1 was recloned as a HindIII-BamHI fragment into a pSV vector, PSV-hyg-HuCK with a hygromycin resistance marker and a human kappa constant domain (FIG. 8 ). The assembly of the genes is summarized inFIG. 9 . - The vectors thus produced were linearized with PvuI (in the case of the pSV-hygro vectors the PvuI digest is only partial) and cotransfected into the non-secreting mouse myeloma line NSO [38] by electroporation [39]. One day after cotransfection, cells were selected in 0.3 μg/ml mycophenolic acid (MPA) and after seven days in 1 μg/ml MPA. After 14 days, four wells, each containing one or two major colonies, were screened by incorporation of 14C-lysine [40] and the secreted antibody detected after precipitation with protein-A Sepharose™ (Pharmacia) on SDS-PAGE [41]. The gels were stained, fixed, soaked in a fluorographic reagent, Amplify™ (Amersham), dried and autoradiographed on preflashed film at −70° C. for 2 days.
- Supernatant was also tested for binding to the mammary carcinoma line MCF-7 and the colon carcinoma line HT-29, essentially as described by Menard et al. [23], either by an indirect immunofluorescence assay on cell suspensions (using a fluorescein-labelled goat anti-human IgG (Amersham)) or by a solid phase RIA on monolayers of fixed cells (using 125I-protein A (Amersham)).
- It was found that one of the supernatants from the four wells contained secreted antibody. The chimeric antibody in the supernatant, like the parent mouse MBr1 antibody, was found to bind to MCF-7 cells but not the HT-29 cells, thus showing that the specificity had been properly cloned and expressed.
- Preparation of DNA from Spleen
- The DNA from the mouse spleen was prepared in one of two ways (although other ways can be used).
-
Method 1. A mouse spleen was cut into two pieces and each piece was put into a standard Eppendorf tube with 200 μl of PBS. The tip of a 1 ml glass pipette was closed and rounded in the blue flame of a Bunsen burner. The pipette was used to squash the spleen piece in each tube. The cells thus produced were transferred to a fresh Eppendorf tube and the method was repeated three times until the connective tissue of the spleen appeared white. Any connective tissue which has been transferred with the cells was removed using a drawn-out Pasteur pipette. The cells were then washed in PBS and distributed into four tubes. - The mouse spleen cells were then sedimented by a 2 minute spin in a Microcentaur centrifuge at low speed setting. All the supernatant was aspirated with a drawn out Pasteur pipette. If desired, at this point the cell sample can be frozen and stored at −20° C.
- To the cell sample (once thawed if it had been frozen) was added 500 μl of water and 5 μl of a 10% solution of NP-40, a non-ionic detergent. The tube was closed and a hole was punched in the lid. The tube was placed on a boiling water bath for 5 minutes to disrupt the cells and was then cooled on ice for 5 minutes. The tube was then spun for 2 minutes at high speed to remove cell debris.
- The supernatant was transferred to a new tube and to this was added 125 μl 5M NaCl and 30 μl 1M MOPS adjusted to pH 7.0. The DNA in the supernatant was absorbed on a
Quiagen 5 tip and purified following the manufacturer's instructions for lambda DNA. After isopropanol precipitation, the DNA was resuspended in 500 μl water. -
Method 2. This method is based on the technique described in Maniatis et al. [30]. A mouse spleen was cut into very fine pieces and put into a 2 ml glass homogenizer. The cells were then freed from the tissue by several slow up and down strokes with the piston. The cell suspension was made in 500 μl phosphate buffered saline (PBS) and transferred to an Eppendorf tube. The cells were then spun for 2 min at low speed in a Microcentaur centrifuge. This results in a visible separation of white and red cells. The white cells, sedimenting slower, form a layer on top of the red cells. The supernatant was carefully removed and spun to ensure that all the white cells had sedimented. The layer of white cells was resuspended in two portions of 500 μl PBS and transferred to another tube. - The white cells were precipitated by spinning in the Microcentaur centrifuge at low speed for one minute. The cells were washed a further two times with 500 μl PBS, and were finally resuspended in 200 μl PBS. The white cells were added to 2.5
ml 25 mM EDTA and 10 mM Tris.Cl, pH 7.4, and vortexed slowly. Whilevortexing 25μl 20% SDS was added. The cells lysed immediately and the solution became viscous and clear. 100 μl of 20 mg/ml proteinase K was added and incubated one to three hours at 50° C. - The sample was extracted with an equal volume of phenol and the same volume of chloroform, and vortexed. After centrifuging, the aqueous phase was removed and 1/10 volume 3M ammonium acetate was added. This was overlaid with three volumes of cold ethanol and the tube rocked carefully until the DNA strands became visible. The DNA was spooled out with a Pasteur pipette, the ethanol allowed to drip off, and the DNA transferred to 1 ml of 10 mM Tris.Cl pH 7.4, 0.1 mM EDTA in an Eppendorf tube. The DNA was allowed to dissolve in the cold overnight on a roller.
- Amplification From Genomic DNA
- The DNA solution was diluted 1/10 in water and boiled for 5 min prior to using the polymerase chain reaction (PCR). For each PCR reaction, typically 50-200 ng of DNA were used.
- The heavy and light chain variable domain encoding sequences in the genomic DNA isolated from the human PBL or the mouse spleen cells was then amplified and cloned using the general protocol described in the first two paragraphs of the section headed “Amplification from RNA/DNA Hybrid” in Example 1, except that during the annealing part of each cycle, the temperature was held at 65° C. and that 30 cycles were used. Furthermore, to minimize the annealing between the 3′ ends of the two primers, the sample was first heated to 95° C., then annealed at 65° C., and only then was the Taq polymerase added. At the end of the 30 cycles, the reaction mixture was held at 60° C. for five minutes to ensure that complete elongation and renaturation of the amplified fragments had taken place.
- The primers used to amplify the mouse spleen genomic DNA were VH1FOR and VH1BACK, for the heavy chain variable domain and VK2FOR and VK1BACK, for the light chain variable domain. (VK2FOR only differs from VK1FOR in that it has an extra C residue on the 5′ end.)
- Other sets of primers, designed to optimize annealing with different families of mouse VH and Vκ genes were devised and used in mixtures with the primers above. For example, mixtures of VK1FOR, MOJK1FOR, MOJK3FOR and MOJK4FOR were used as forward primers and mixtures of VK1BACK, MOVKIIABACK and MOVKIIBBACK as back primers for amplification of Vκ genes. Likewise mixtures of VH1FOR, MOJH1FOR, MOJH2FOR, MOJH3FOR and MOJH4FOR were used as forward primers and mixtures of VH1BACK, MOVHIBACK, MOVHIIABACK, MOVHIIBBACK, MOVHIIIBACK were used as backward primers for amplification of VH genes.
- All these heavy chain FOR primers referred to above contain a BstEII site and all the BACK primers referred to above contain a PstI site. These light chain FOR and BACK primers referred to above all contain BglII and PvuII sites respectively. Light chain primers (VK3FOR and VK2BACK) were also devised which utilized different restriction sites, SacI and XhoI.
- Typically all these primers yielded amplified DNA of the correct size on gel electrophoresis, although other bands were also present. However, a problem was identified in which the 5′ and 3′ ends of the forward and backward primers for the VH genes were partially complementary, and this could yield a major band of “primer-dimer” in which the two oligonucleotides prime on each other. For this reason an improved forward primer, VH1FOR-2 was devised in which the two 3′ nucleotides were removed from VH1FOR.
- Thus, the preferred amplification conditions for mouse VH genes are as follows: the sample was made in a volume of 50-100 μl, 50-100 ng of DNA, VH1FOR-2 and VH1BACK primers (25 pmole of each), 250 μM of each deoxynucleotide triphosphate, 10 mM Tris.HCl, pH 8.8, 50 mM KCl, 1.5 mM MgCl2, and 100 μg/ml gelatine. The sample was overlaid with paraffin oil, heated to 95° C. for 2 min, 65° C. for 2 min, and then to 72° C.: taq polymerase was added after the sample had reached the elongation temperature and the reaction continued for 2 min at 72° C. The sample was subjected to a further 29 rounds of temperature cycling using the Techne PHC-1 programmable heating block.
- The preferred amplification conditions for mouse Vκ genes from genomic DNA are as follows: the sample treated as above except with Vκ primers, for example VK3FOR and VK2BACK, and using a cycle of 94° C. for one minute, 60° C. for one minute and 72° C. for one minute.
- The conditions which were devised for genomic DNA are also suitable for amplification from the cDNA derived from mRNA from mouse spleen or mouse hybridoma.
- Cloning and Analysis of Variable Region Genes
- The reaction mixture was then extracted twice with 40 μl of water-saturated diethyl ether. This was followed by a standard phenol extraction and ethanol precipitation as described in Example 1. The DNA pellet was then dissolved in 100
μl 10 mM Tris.Cl, 0.1 mM EDTA. - Each reaction mixture containing a light chain variable domain encoding sequence was digested with SacI and XhoI (or with PvuII and BglII) to enable it to be ligated into a suitable expression vector. Each reaction mixture containing a heavy chain variable domain encoding sequence was digested with PstI and BstEII for the same purpose.
- The heavy chain variable genes isolated as above from a mouse hyper-immunized with lysozyme were cloned into M13VHPCR1 vector and sequenced. The complete sequences of 48 VH gene clones were determined (
FIGS. 10 a-10 b). All but two of the mouse VH gene families were represented, with frequencies of: VA (1), IIIC (1), IIIB (8), IIIA (3), IIB (17), IIA (2), IB (12), IA (4). In 30 clones, the D segments could be assigned to families SP2 (14), FL16 (11) and Q52 (5), and in 38 clones the JH minigenes to families JH1 (3), JH2 (7), JH3 (14) and JH4 (14). The different sequences of CDR3 marked out each of the 48 clones as unique. Nine pseudogenes and 16 unproductive rearrangements were identified. Of the clones sequenced, 27 have open reading frames. - Thus the method is capable of generating a diverse repertoire of heavy chain variable genes from mouse spleen DNA.
- Preparation of mRNA
- Human peripheral blood lymphocytes were purified and mRNA prepared directly (Method 1), or mRNA was prepared after addition of Epstein Barr virus (Method 2).
-
Method 1. 20 ml of heparinized human blood from a healthy volunteer was diluted with an equal volume of phosphate buffered saline (PBS) and distributed equally into 50 ml Falcon tubes. The blood was then underlayed with 15 ml Ficoll Hypaque (Pharmacia 10-A-001-07). To separate the lymphocytes from the red blood cells, the tubes were spun for 10 minutes at 1800 rpm at room temperature in an IEC Centra 3E table centrifuge. The peripheral blood lymphocytes (PBL) were then collected from the interphase by aspiration with a Pasteur pipette. The cells were diluted with an equal volume of PBS and spun again at 1500 rpm for 15 minutes. The supernatant was aspirated, the cell pellet was resuspended in 1 ml PBS and the cells were distributed into two Eppendorf tubes. -
Method 2. 40 ml human blood from a patient with HIV in the pre-AIDS condition was layered on Ficoll to separate the white cells (seeMethod 1 above). The white cells were then incubated in tissue culture medium for 4-5 days. Onday 3, they were infected with Epstein Barr virus. The cells were pelleted (approx 2×107 cells) and washed in PBS. - The cells were pelleted again and lysed with 7 ml 5M guanidine isothiocyanate, 50 mM Tris, 10 mM EDTA, 0.1 mM dithiothreitol. The cells were vortexed vigorously and 7 volumes of 4M LiCl added. The mixture was incubated at 4° C. for 15-20 hrs. The suspension was spun and the supernatant resuspended in 3M LiCl and centrifuged again. The pellet was dissolved in 2 ml 0.1% SDS, 10 mM Tris HCl and 1 mM EDTA. The suspension was frozen at −20° C., and thawed by vortexing for 20 s every 10 min for 45 min. A large white pellet was left behind and the clear supernatant was extracted with phenol chloroform, then with chloroform. The RNA was precipitated by adding 1/10 volume 3M sodium acetate and 2 vol ethanol and leaving overnight at −20° C. The pellet was suspended in 0.2 ml water and reprecipitated with ethanol. Aliquots for cDNA synthesis were taken from the ethanol precipitate which had been vortexed to create a fine suspension.
- 100 μl of the suspension was precipitated and dissolved in 20 μl water for cDNA synthesis [30] using 10 pmole of a HUFOR primer (see below) in final volume of 50 μl. A sample of 5 μl of the cDNA was amplified as in Example 2 except using the primers for the human VH gene families (see below) using a cycle of 95° C., 60° C. and 72° C.
- The back primers for the amplification of human DNA were designed to match the available human heavy and light chain sequences, in which the different families have slightly different nucleotide sequences at the 5′ end. Thus for the human VH genes, the primers Hu2VHIBACK, HuVHIIBACK, Hu2VHIIIBACK and HuVH1VBACK were designed as back primers, and HUJH1FOR, HUJH2FOR and HUJH4FOR as forward primers based entirely in the variable gene. Another set of forward primers Hu1VHFOR, Hu2VHFOR, Hu3VHFOR, and Hu4VHFOR was also used, which were designed to match the human J-regions and the 5′ end of the constant regions of different human isotypes.
- Using sets of these primers it was possible to demonstrate a band of amplified ds cDNA by gel electrophoresis.
- One such experiment was analyzed in detail to establish whether there was a diverse repertoire in a patient with HIV infection. It is known that during the course of AIDS, that T-cells and also antibodies are greatly diminished in the blood. Presumably the repertoire of lymphocytes is also diminished. In this experiment, for the forward priming, an equimolar mixture of primers Hu1VHFOR, Hu2VHFOR, Hu3VHFOR, and Hu4VHFOR (in
PCR 25 pmole ofprimer 5′ ends) was used. For the back priming, the primers Hu2VHIBACK, HuVHIIBACK, Hu2VHIIIBACK and HuVH1VBACK were used separately in four separate primings. The amplified DNA from the separate primings was then pooled, digested with restriction enzymes PstI and BstEII as above, and then cloned into the vector M13VHPCR1 for sequencing. The sequences reveal a diverse repertoire (FIG. 11 ) at this stage of the disease. - For human Vκ genes the primers HuJK1FOR, HUJK3FOR; HUJK4FOR and HUJK5FOR were used as forward primers and VK1BACK as back primer. Using these primers it was possible to see a band of amplified ds cDNA of the correct size by gel electrophoresis.
- Human peripheral blood lymphocytes of a patient with non-Hodgkins lymphoma were prepared as in Example 3 (Method 1). The genomic DNA was prepared from the PBL using the technique described in Example 2 (Method 2). The VH region in the isolated genomic DNA was then amplified and cloned using the general protocol described in the first two paragraphs of the section headed “Amplification from RNA/DNA hybrid” in Example 1 above, except that during the annealing part of each cycle, the temperature was held at 55° C. and that 30 cycles were used. At the end of the 30 cycles, the reaction mixture was held at 60° C. for five minutes to ensure that complete elongation and renaturation of the amplified fragments had taken place.
- The forward primer used was HuHep1FOR, which contains a SacI site. This primer is designed to anneal to the 3′ end of the unrearranged human VH region gene, and in particular includes a sequence complementary to the last three codons in the VH region gene and nine nucleotides downstream of these three codons.
- As the back primer, an equimolar mixture of HuOcta1BACK, HuOcta2BACK and HuOcta3BACK was used. These primers anneal to a sequence in the promoter region of the genomic DNA VH gene (see
FIG. 1 ). 5 μl of the amplified DNA was checked on 2% agarose gels in TBE buffer and stained with ethidium bromide. A double band was seen of about 620 nucleotides which corresponds to the size expected for the unrearranged VH gene. The ds cDNA was digested with SacI and cloned into an M13 vector for sequencing. Although there are some sequences which are identical, a range of different unrearranged human VH genes were identified (FIG. 12 ). - The heavy chain variable domain (VHLYS) of the D1.3 (anti-lysozyme) antibody was cloned into a vector similar to that described previously [42] but under the control of the lac z promoter, such that the VHLYS domain is attached to a pelB leader sequence for export into the periplasm. The vector was constructed by synthesis of the pelB leader sequence [43], using overlapping oligonucleotides, and cloning into a pUC 19 vector [35]. The VHLYS domain of the D1.3 antibody was derived from a cDNA clone [44] and the construct (PSW1) sequenced (
FIG. 13 ). - To express both heavy and light chain variable domains together, the light chain variable region (VKLYS) of the D1.3 antibody was introduced into the pSW1 vector, with a pelB signal sequence to give the construct pSW2 (
FIGS. 14 a-14 b). - A strain of E. coli (BMH71-18) [45] was then transformed [46,47] with the plasmid pSW1 or pSW2, and colonies resistant to ampicillin (100 μg/ml) were selected on a rich (2×TY=per litre of water, 16 g Bacto-tryptone, 10 g yeast extract, 5 g NaCl) plate which contained 1% glucose to repress the expression of variable domain(s) by catabolite repression.
- The colonies were inoculated into 50
ml 2×TY (with 1% glucose and 100 μg/ml ampicillin) and grown in flasks at 37° C. with shaking for 12-16 hr. The cells were centrifuged, the pellet washed twice with 50 mM sodium chloride, resuspended in 2×TY medium containing 100 μg/ml ampicillin and the inducer IPTG (1 mM) and grown for a further 30 hrs at 37° C. The cells were centrifuged and the supernatant was passed through a Nalgene filter (0.45 μm) and then down a 1-5 ml lysozyme-Sepharose® affinity column (Pharmacia Fine Chemicals, Inc.). (The column was derived by coupling lysozyme at 10 mg/ml to CNBr activated Sepharose.) The column was first washed with phosphate buffered saline (PBS), then with 50 mM diethylamine to elute the VHLYS domain (from pSW1) or VHLYS in association with VKLYS (from pSW2). - The VHLYS and VKLYS domains were identified by SDS polyacrylamide electrophoresis as the correct size. In addition, N-terminal sequence determination of VHLYS and VKLYS isolated from a polyacrylamide gel showed that the signal peptide had been produced correctly. Thus both the Fv fragment and the VHLYS domains are able to bind to the lysozyme affinity column, suggesting that both retain at least some of the affinity of the original antibody.
- The size of the VHLYS domain was compared by FPLC with that of the Fv fragment on Superose 12. This indicates that the VHLYS domain is a monomer. The binding of the VHLYS and Fv fragment to lysozyme was checked by ELISA, and equilibrium and rapid reaction studies were carried out using fluorescence quench.
- The ELISA for lysozyme binding was undertaken as follows:
- (1) The plates (Dynatech Immulon) were coated with 200 μl per well of 300 μg/ml lysozyme in 50 mM NaHCO3, pH 9.6 overnight at room temperature;
- (2) The wells were rinsed with three washes of PBS, and blocked with 300 μl per well of 1% Sainsbury's instant dried skimmed milk powder in PBS for 2 hours at 37° C.;
- (3) The wells were rinsed with three washes of PBS and 200 μl of VHLYS or Fv fragment (VHLYS associated with VKLYS) were added and incubated for 2 hours at room temperature;
- (4) The wells were washed three times with 0.05
% Tween 20 in PBS and then three times with PBS to remove detergent; - (5) 200 μl of a suitable dilution (1:1000) of rabbit polyclonal antisera raised against the Fv fragment in 2% skimmed milk powder in PBS was added to each well and incubated at room temperature for 2 hours;
- (6) Washes were repeated as in (4);
- (7) 200 μl of a suitable dilution (1:1000) of goat anti-rabbit antibody (ICN Immunochemicals) coupled to horse radish peroxidase, in 2% skimmed milk powder in PBS, was added to each well and incubated at room temperature for 1 hour;
- (8) Washes were repeated as in (4); and
- (9) 200
ml μl 20% hydrogen peroxide: water per 10 ml) was added to each well and the color allowed to develop for up to 10 minutes at room temperature. - The reaction was stopped by adding 0.05% sodium azide in 50 mM citric acid pH 4.3. ELISA plates were read in a Titertek Multiscan plate reader. Supernatant from the induced bacterial cultures of both pSW1 (VHLYS domain) or pSW2 (Fv fragment) was found to bind to lysozyme in the ELISA.
- The purified VHLYS and Fv fragments were titrated with lysozyme using fluorescence quench (Perkin Elmer LS5B Luminescence Spectrometer) to measure the stoichiometry of binding and the affinity constant for lysozyme [48,49]. The titration of the Fv fragment at a concentration of 30 nM indicates a dissociation constant of 2.8 nM using a Scatchard analysis.
- A similar analysis using fluorescence quench and a Scatchard plot was carried out for VHLYS, at a VHLYS concentration of 100 nM. The stoichiometry of antigen binding is about 1 mole of lysozyme per mole of VHLYS (calculated from plot). (The concentration of VH domains was calculated from optical density at 280 nM using the typical extinction coefficient for complete immunoglobulins.) Due to possible errors in measuring low optical densities and the assumption about the extinction coefficient, the stoichiometry was also measured more carefully. VHLYS was titrated with lysozyme as above using fluorescence quench. To determine the concentration of VHLYS a sample of the stock solution was removed, a known amount of norleucine added, and the sample subjected to quantitative amino acid analysis. This showed a stoichiometry of 1.2 mole of lysozyme per mole of VHLYS domain. The dissociation constant was calculated as about 12 nM.
- The on-rates for VHLYS and Fv fragments with lysozyme were determined by stopped-flow analysis (HI Tech Stop Flow SHU machine) under pseudo-first order conditions with the fragment at a ten fold higher concentration than lysozyme [50]. The concentration of lysozyme binding sites was first measured by titration with lysozyme using fluorescence quench as above. The on rates were calculated per mole of binding site (rather than amount of VHLYS protein). The on-rate for the Fv fragment was found to be 2.2×106 M−1 s−1 at 25° C. The on-rate for the VHLYS fragment found to be 3.8×106 M−1 s−1 and the off-rate 0.075 s−1 at 20° C. The calculated affinity constant is 19 nM. Thus the VHLYS binds to lysozyme with a dissociation constant of about 19 nM, compared with that of the Fv of 3 nM.
- A mouse was immunized with hen egg white lysozyme (100 μg i.p.
day 1 in complete Freunds adjuvant), after 14 days immunized i.p. again with 100 μg lysozyme with incomplete Freunds adjuvant, and onday 35 i.v. with 50 μg lysozyme in saline. On day 39, spleen was harvested. A second mouse was immunized with keyhole limpet hemocyanin (KLH) in a similar way. The DNA was prepared from the spleen according to Example 2 (Method 2). The VH genes were amplified according to the preferred method in Example 2. - Human peripheral blood lymphocytes from a patient infected with HIV were prepared as in Example 3 (Method 2) and mRNA prepared. The VH genes were amplified according to the method described in Example 3, using primers designed for human VH gene families.
- After the PCR, the reaction mixture and oil were extracted twice with ether, once with phenol and once with phenol/CHCl3. The double stranded DNA was then taken up in 50 μl of water and frozen. 5 μl was digested with PstI and BstEII (encoded within the amplification primers) and loaded on an agarose gel for electrophoresis. The band of amplified DNA at about 350 bp was extracted.
- Expression of Anti-Lysozyme Activities
- The repertoire of amplified heavy chain variable domains (from mouse immunized with lysozyme and from human PBLs) was then cloned directly into the expression vector pSW1HPOLYMYC. This vector is derived from pSW1 except that the VHLYS gene has been removed and replaced by a polylinker restriction site. A sequence encoding a peptide tag was inserted (
FIG. 15 ). Colonies were toothpicked into 1 ml cultures. After induction (see Example 5 for details), 10 μl of the supernatant from fourteen 1 ml cultures was loaded on SDS-PAGE gels and the proteins transferred electrophoretically to nitrocellulose. The blot was probed with antibody 9E10 directed against the peptide tag. - The probing was undertaken as follows. The nitrocellulose filter was incubated in 3% bovine serum albumin (BSA)/TBS buffer for 20 min (10×TBS buffer is 100 mM Tris.HCl, pH 7.4, 9% w/v NaCl). The filter was incubated in a suitable dilution of antibody 9E10 (about 1/500) in 3% BSA/TBS for 1-4 hrs. After three washes in TBS (100 ml per wash, each wash for 10 min), the filter was incubated with 1:500 dilution of anti-mouse antibody (peroxidase conjugated anti-mouse Ig (Dakopats)) in 3% BSA/TBS for 1-2 hrs. After three washes in TBS and 0.1% Triton X-100 (about 100 ml per wash, each wash for 10 min), a solution containing 10 ml chloronaphthol in methanol (3 mg/ml), 40 ml TBS and 50 μl hydrogen peroxide solution was added over the blot and allowed to react for up to 10 min. The substrate was washed out with excess water. The blot revealed bands similar in mobility to VHLYSMYC on the Western blot, showing that other VH domains could be expressed.
- Colonies were then toothpicked individually into wells of an ELISA plate (200 μl) for growth and induction. They were assayed for lysozyme binding with the 9E10 antibody (as in Examples 5 and 7). Wells with lysozyme-binding activity were identified. Two positive wells (of 200) were identified from the amplified mouse spleen DNA and one well from the human cDNA. The heavy chain variable domains were purified on a column of lysozyme-Sepharose. The affinity for lysozyme of the clones was estimated by fluorescence quench titration as >50 nM. The affinities of the two clones (VH3 and VH8) derived from the mouse genes were also estimated by stop flow analysis (ratio of koff/kon) as 12 nM and 27 nM respectively. Thus both these clones have a comparable affinity to the VHLYS domain. The encoded amino acid sequences of VH3 and VH8 are given in
FIG. 16 , and that of the human variable domain inFIG. 17 . - A library of VH domains made from the mouse immunized with lysozyme was screened for both lysozyme and keyhole limpet hemocyanin (KLH) binding activities. Two thousand colonies were toothpicked in groups of five into wells of ELISA plates, and the supernatants tested for binding to lysozyme coated plates and separately to KLH coated plates. Twenty one supernatants were shown to have lysozyme binding activities and two to have KLH binding activities. A second expression library, prepared from a mouse immunized with KLH was screened as above. Fourteen supernatants had KLH binding activities and a single supernatant had lysozyme binding activity.
- This shows that antigen binding activities can be prepared from single VH domains, and that immunization facilitates the isolation of these domains.
- Taking a single rearranged VH gene, it may be possible to derive entirely new antigen binding activities by extensively mutating each of the CDRs. The mutagenesis might be entirely random, or be derived from pre-existing repertoires of CDRs. Thus a repertoire of CDR3s might be prepared as in the preceding examples by using “universal” primers based in the flanking sequences, and likewise repertoires of the other CDRs (singly or in combination). The CDR repertoires could be stitched into place in the flanking framework regions by a variety of recombinant DNA techniques.
- CDR3 appears to be the most promising region for mutagenesis as CDR3 is more variable in size and sequence than
CDRs - Multiple mutations were made in CDR3. The polymerase chain reaction (PCR) and a highly degenerate primer were used to make the mutations and by this means the original sequence of CDR3 was destroyed. (It would also have been possible to construct the mutations in CDR3 by cloning a mixed oligonucleotide duplex into restriction sites flanking the CDR or by other methods of site-directed mutagenesis). Mutants expressing heavy chain variable domains with affinities for lysozyme were screened and those with improved affinities or new specificities were identified.
- The source of the heavy chain variable domain was an M113 vector containing the VHLYS gene. The body of the sequence encoding the variable region was amplified using the polymerase chain reaction (PCR) with the mutagenic primer VHMUT1 based in CDR3 and the M13 primer which is based in the M13 vector backbone. The mutagenic primer hypermutates the central four residues of CDR3 (Arg-Asp-Tyr-Arg). The PCR was carried out for 25 cycles on a Techne PHC-1 programmable heat block using 100 ng single stranded M13 mp19SWO template, with 25 pmol of VHMUT1 and the M13 primer, 0.5 mM each dNTP, 67 mM Tris.HCl, pH 8.8, 10 mM MgCl2, 17 mM (NH4)2SO4, 200 μg/ml gelatine and 2.5 units Taq polymerase in a final volume of 50 μl. The temperature regime was 95° C. for 1.5 min, 25° C. for 1.5 min and 72° C. for 3 min (However a range of PCR conditions could be used). The reaction products were extracted with phenol/chloroform, precipitated with ethanol and resuspended in 10 mM Tris. HCl and 0.1 mM EDTA, pH 8.0.
- The products from the PCR were digested with PstI and BstEII and purified on a 1.5% LGT agarose gel in Tris acetate buffer using Geneclean® (Bio 101, LaJolla). The gel purified band was ligated into pSW2HPOLY (
FIG. 19 ). (This vector is related to pSW2 except that the body of the VHLYS gene has been replaced by a polylinker.) The vector was first digested with BstEII and PstI and treated with calf-intestinal phosphatase. Aliquots of the reaction mix were used to transform E. coli BMH 71-18 to ampicillin resistance. Colonies were selected on ampicillin (100 μg/ml) rich plates containing glucose at 0.8% w/v. - Colonies resulting from transfection were picked in pools of five into two 96 well Corning microtitre plates, containing 200
μl 2×TY medium and 100 μl TY medium, 100 μg/ml ampicillin and 1% glucose. The colonies were grown for 24 hours at 37° C. and then cells were washed twice in 200μl 50 mM NaCl, pelleting the cells in an IEC Centra-3 bench top centrifuge with microtitre plate head fitting. Plates were spun at 2,500 rpm for 10 min at room temperature. Cells were resuspended in 200μl 2×TY, 100 μg/ml ampicillin and 1 mM IPTG (Sigma) to induce expression, and grown for a further 24 hr. - Cells were spun down and the supernatants used in ELISA with lysozyme coated plates and anti-idiotypic sera (raised in rabbits against the Fv fragment of the D1.3 antibody). Bound anti-idiotypic serum was detected using horse radish peroxidase conjugated to anti-rabbit sera (ICN Immunochemicals). Seven of the wells gave a positive result in the ELISA. These pools were restreaked for single colonies which were picked, grown up, induced in microtitre plates and rescreened in the ELISA as above. Positive clones were grown up at the 50 ml scale and expression was induced. Culture supernatants were purified as in Example 5 on columns of lysozyme-Sepharose and eluates analysed on SDS-PAGE and staining with Page Blue 90 (BDH). On elution of the column with diethylamine, bands corresponding to the VHLYS mutant domains were identified, but none to the VKLYS domains. This suggested that although the mutant domains could bind to lysozyme, they could no longer associate with the VKYLS domains.
- For seven clones giving a positive reaction in ELISA, plasmids were prepared and the VKLYS gene excised by cutting with EcoRI and religating. Thus the plasmids should only direct the expression of the VHLYS mutants. 1.5 ml cultures were grown and induced for expression as above. The cells were spun down and supernatant shown to bind lysozyme as above. (Alternatively the amplified mutant VKLYS genes could have been cloned directly into the pSW1HPOLY vector for expression of the mutant activities in the absence of VKLYS.)
- An ELISA method was devised in which the activities of bacterial supernatants for binding of lysozyme (or KLH) were compared. Firstly a vector was devised for tagging of the VH domains at its C-terminal region with a peptide from the c-myc protein which is recognized by a monoclonal antibody 9E10. The vector was derived from pSW1 by a BstEII and SmaI double digest, and ligation of an oligonucleotide duplex made from
-
5′ GTC ACC GTC TCC TCA GAA CAA AAA CTC ATA TCA GAA GAG GAT CTG AAT TAA TAA 3′and 5′ TTA TTA ATT CAG ATC CTC TTC TGA GAT GAG TTT TTG TTC TGA GGA GAC G 3′. - The VHLYSMYC protein domain expressed after induction was shown to bind to lysozyme and to the 9E10 antibody by ELISA as follows:
- (1) Falcon (3912) flat bottomed wells were coated with 180 μl lysozyme (3 mg/ml) or KLH (50 μg/ml) per well in 50 mM NaHCO3, pH 9.6, and left to stand at room temperature overnight;
- (2) The wells were washed with PBS and blocked for 2 hrs at 37° C. with 200
μl 2% Sainsbury's instant dried skimmed milk powder in PBS per well; - (3) The Blocking solution was discarded, and the walls washed out with PBS (3 washes) and 150 μl test solution (supernatant or purified tagged domain) pipetted into each well. The sample was incubated at 37° C. for 2 hrs;
- (4) The test solution was discarded, and the wells washed out with PBS (3 washes). 100 μl of 4 μg/ml purified 9E10 antibody in 2% Sainsbury's instant dried skimmed milk powder in PBS was added, and incubated at 37° C. for 2 hrs;
- (5) The 9E10 antibody was discarded, the wells washed with PBS (3 washes). 100 ml of 1/500 dilution of anti-mouse antibody (peroxidase conjugated anti-mouse Ig (Dakopats)) was added and incubated at 37° C. for 2 hrs;
- (6) The second antibody was discarded and wells washed three times with PBS; and
- (7) 100
μl μl 20% hydrogen peroxide: water per 10 ml) was added to each well and the color allowed to develop for up to 10 minutes at room temperature. - The reaction was stopped by adding 0.05% sodium azide in 50 mM citric acid, pH 4.3. ELISA plates were read in an Titertek Multiscan plate reader.
- The activities of the mutant supernatants were compared with VHLYS supernatant by competition with the VHLYSMYC domain for binding to lysozyme. The results show that supernatant from clone VHLYSMUT59 is more effective than wild type VHLYS supernatant in competing for VHLYSMYC. Furthermore, Western blots of SDS-PAGE aliquots of supernatant from the VHLYS and VHLYSMUT59 domain (using anti-Fv antisera) indicated comparable amounts of the two samples. Thus assuming identical amounts of VHLYS and VHLYSMUT59, the affinity of the mutant appears to be greater than that of the VHLYS domain.
- To check the affinity of the VHLYSMUT59 domain directly, the clone was grown at the 1 L scale and 200-300 μg purified on lysozyme-Sepharose as in Example 5. By fluorescence quench titration of samples of VHLYS and VHLYSMUT59, the number of binding sites for lysozyme were determined. The samples of VHLYS and VHLYSMUT59 were then compared in the competition ELISA with VHLYSMYC over two orders of magnitude. In the competition assay each microtitre well contained a constant amount of VHLYSMYC (approximately 0.6 μg VHLYSMYC). Varying amounts of VHLYS or VHLYSMUT59 (3.8 μM in lysozyme binding sites) were added (0.166-25 μl). The final volume and buffer concentration in all wells was constant. 9E10 (anti-myc) antibody was used to quantitate bound VHLYSMYC in each assay well. The % inhibition of VHLYSMYC binding was calculated for each addition of VHLYS or VHLYSMUT59, after subtraction of background binding. Assays were carried out in duplicate. The results indicate that VHLYSMUT59 has a higher affinity for lysozyme than VHLYS.
- The VHLYSMUT59 gene was sequenced (after recloning into M13) and shown to be identical to the VHLYS gene except for the central residues of CDR3 (Arg-Asp-Tyr-Arg). These were replaced by Thr-Gln-Arg-Pro: (encoded by ACACAAAGGCCA).
- A library of 2000 mutant VH clones was screened for lysozyme and also for KLH binding (
toothpicking 5 colonies per well as described in Example 6). Nineteen supernatants were identified with lysozyme binding activities and four with KLH binding activities. This indicates that new specificities and improved affinities can be derived by making a random repertoire of CDR3. - The finding that single domains have excellent binding activities should allow the construction of strings of domains (concatamers). Thus, multiple specificities could be built into the same molecule, allowing binding to different epitopes spaced apart by the distance between domain heads. Flexible linker regions could be built to space out the domains. In principle such molecules could be devised to have exceptional specificity and affinity.
- Two copies of the cloned heavy chain variable gene of the D1.3 antibody were linked by a nucleotide sequence encoding a flexible linker Gly-Gly-Gly-Ala-Pro-Ala-Ala-Ala-Pro-Ala-Gly-Gly-Gly- (by several steps of cutting, pasting and site directed mutagenesis) to yield the plasmid pSW3 (
FIG. 20 ). The expression was driven by a lacZ promoter and the protein was secreted into the periplasm via a pelB leader sequence (as described in Example 5 for expression of pSW1 and pSW2). The protein could be purified to homogeneity on a lysozyme affinity column. On SDS polyacrylamide gels, it gave a band of the right size (molecular weight about 26,000). The protein also bound strongly to lysozyme as detected by ELISA (see Example 5) using anti-idiotypic antiserum directed against the Fv fragment of the D1.3 antibody to detect the protein. Thus, such constructs are readily made and secreted and at least one of the domains binds to lysozyme. - A cysteine residue was introduced at the C-terminus of the VHLYS domain in the vector pSW2. The cysteine was introduced by cleavage of the vector with the restriction enzymes BstI and SmaI (which excises the C-terminal portion of the J segment) and ligation of a short oligonucleotide duplex
-
5′ GTC ACC GTC TCC TCA TGT TAA TAA 3′and 5′ TTA TTA ACA TGA GGA GAC G 3′.
By purification on an affinity column of lysozyme Sepharose it was shown that the VHLYS-Cys domain was expressed in association with the VKLYS variable domain, but the overall yields were much lower than the wild type Fv fragment. Comparison of non-reducing and reducing SDS polyacrylamide gels of the purified Fv-Cys protein indicated that the two VH-Cys domains had become linked-through the introduced cysteine residue. - Linking of enzyme activities to VH domains should be possible by either cloning the enzyme on either the N-terminal or the C-terminal side of the VH domain. Since both partners must be active, it may be necessary to design a suitable linker (see Example 8) between the two domains. For secretion of the VH-enzyme fusion, it would be preferable to utilize an enzyme which is usually secreted. In
FIGS. 21 a-21 c, there is shown the sequence of a fusion of a VH domain with alkaline phosphatase. The alkaline phosphatase gene was cloned from a plasmid carrying the E. coli alkaline phosphatase gene in a plasmid pEK48 [51] using the polymerase chain reaction. The gene was amplified with the primers -
5′CAC CAC GGT CAC CGT CTC CTC ACG GAC ACC AGA AAT GCC TGT TCT G 3′and 5′ GCG AAA ATT CAC TCC CGG GCG CGG TTT TAT TTC 3′.
The gene was introduced into the vector pSW1 by cutting at BstEII and SmaI. The construction (FIGS. 21 a-21 c) was expressed in E. coli strain BMH71-18 as in Example 5 and screened for phosphatase activity using 1 mg/ml p-nitrophenylphosphate as substrate in 10 mM diethanolamine and 0.5 mM MgCl2, pH 9.5) and also on SDS polyacrylamide gels which had been Western blotted (detecting with anti-idiotypic antiserum). No evidence was found for the secretion of the linked VHLYS-alkaline phosphatase as detected by Western blots (see Example 5), or for secretion of phosphatase activity. - However when the construct was transfected into a bacterial strain BL21DE3 [52] which is deficient in proteases, a band of the correct size (as well as degraded products) was detected on the Western blots. Furthermore phosphatase activity could now be detected in the bacterial supernatant. Such activity is not present in supernatant from the strain which had not been transfected with the construct.
- A variety of linker sequences could then be introduced at the BstEII site to improve the spacing between the two domains.
- A repertoire of Vκ genes was derived by PCR using primers as described in Example 2 from DNA prepared from mouse spleen and also from mouse spleen mRNA using the primers VK3FOR and VK2BACK and a cycle of 94° C. for 1 min, 60° C. for 1 min, 72° C. for 2 min. The PCR amplified DNA was fractionated on the agarose gel, the band excised and cloned into a vector which carries the VHLYS domain (from the D 1.3 antibody), and a cloning site (SacI and XhoI) for cloning of the light chain variable domains with a myc tail (pSW1VHLYS-VKPOLYMYC,
FIG. 22 ). - Clones were screened for lysozyme binding activities as described in Examples 5 and 7 via the myc tag on the light chain variable domain, as this should permit the following kinds of Vκ domains to be identified:
- (1) those which bind to lysozyme in the absence of the VHLYS domain;
- (2) those which associate with the heavy chain and make no contribution to binding of lysozyme; and
- (3) those which associate with the heavy chain and also contribute to binding of lysozyme (either helping or hindering).
- This would not identify those Vκ domains which associated with the VHLYS domain and completely abolished its binding to lysozyme.
- In a further experiment, the VHLYS domain was replaced by the heavy chain variable domain VH3 which had been isolated from the repertoire (see Example 6), and then the Vκ domains cloned into the vector. (Note that the VH3 domain has an internal SacI site and this was first removed to allow the cloning of the Vκ repertoire as SacI-XhoI fragments.)
- By screening the supernatant using the ELISA described in Example 6, bacterial supernatants will be identified which bind lysozyme.
- By screening several clones from a VH library derived from a mouse immunized with lysozyme via a Western blot, using the 9E10 antibody directed against the peptide tag, one clone was noted with very high levels of expression of the domain (estimated as 25-50 mg/l). The clone was sequenced to determine the nature of the sequence. The sequence proved to be closely related to that of the VHLYS domain, except with a few amino acid changes (
FIG. 23 ). The result was unexpected, and shows that a limited number of amino acid changes, perhaps even a single amino acid substitution, can cause greatly elevated levels of expression. - By making mutations of the high expressing domain at these residues, it was found that a single amino acid change in the VHLYS domain (
Asn 35 to H is) is sufficient to cause the domain to be expressed at high levels. - It can thus be seen that the present invention enables the cloning, amplification and expression of heavy and light chain variable domain encoding sequences in a much more simple manner than was previously possible. It also shows that isolated variable domains or such domains linked to effector molecules are unexpectedly useful.
- It will be appreciated that the present invention has been described above by way of example only and that variations and modifications may be made by the skilled person without departing from the scope of the invention.
-
- [1] Inbar et al., PNAS-USA, 69, 2659-2662, 1972.
- [2] Amit et al., Science, 233, 747, 1986.
- [3] Satow et al., J. Mol. Biol., 190, 593, 1986.
- [4] Colman et al., Nature, 326, 358, 1987.
- [5] Sheriff et al., PNAS-USA, 84, 8075-8079, 1987.
- [6] Padlan et al., PNAS-USA, 86, 5938-5942, 1989.
- [7] Skerra and Plückthun, Science, 240, 1038-1041, 1988.
- [8] Bird et al., Science, 242, 423-426, 1988.
- [9] Huston et al., PNAS-USA, 85, 5879-5833, 1988.
- [10] Fleischman, Arch. Biochem. Biophys. Suppl., 1, 174, 1966.
- [1,1] Porter and Weir, J. Cell. Physiol. Suppl., 1, 51, 1967.
- [1,2] Jaton et al., Biochemistry, 7, 4185, 1968.
- [1,3] Rockey, J. Exp. Med., 125, 249, 1967.
- [1,4] Stevenson, Biochem. J., 133, 827-836, 1973.
- [15] Edmundson et al., Biochemistry, 14, 3953, 1975.
- [1,6] Rossman et al., Nature, 317, 145-153, 1985.
- [1,7] Saiki et al., Science, 230, 1350-1354, 1985.
- [1,8] Larrick et al., Biochem. Biophys. Res. Comm., 160, 1250, 1989.
- [1,9] Orlandi et al., PNAS-USA, 86, 3833, 1989.
- [20] Yon and Fried, Nuc. Acids Res., 17, 4895, 1989.
- [21] Fields and Song, Nature, 340, 245-246, 1989.
- [22] Baldwin and Schultz, Science, 245, 1104-1107, 1989.
- [23] Menard et al., Cancer Res., 43, 1295-1300, 1983.
- [24] Bosslet et al., Eur. J. Nuc. Med., 14, 523-528, 1988.
- [25] Bosslet et al., Cancer Immunol. Immunother., 23, 185-191, 1986.
- [26] Bosslet et al., Int. J. Cancer, 36, 75-84, 1985.
- [27]
- [28] Bremer et al., J. Biol. Chem., 259, 14773-14777, 1984.
- [29] Griffiths & Milstein, Hybridoma Technology in the Biosciences and Medicine, 103-115, 1985.
- [30] Maniatis et al., Molecular Cloning: a Laboratory Manual, Cold Spring Harbour Laboratory, 1982.
- [31] Jones et al., Nature, 321, 522-525, 1986.
- [32] Zoller & Smith, Nuc. Acids Res., 10, 6457-6500, 1982.
- [33] Carter et al., Nuc. Acids Res., 13, 4431-4443, 1985.
- [34] Sanger et al., PNAS-USA, 74, 5463-5467, 1977.
- [35] Yannisch-Perron et al., Gene, 33, 103-119, 1985.
- [36]
- [37] Riechmann et al., Nature, 332, 323-327, 1988.
- [38] Kearney et al., J. Immunol., 123, 1548-1550, 1979.
- [39] Patter et al., PNAS-USA, 81, 7161-7163, 1984.
- [40] Galfre & Milstein, Meth. Enzym., 73, 1-46, 1981.
- [41] Laemmli, Nature, 227, 680-685, 1970.
- [42] Better et al., Science, 240, 1041, 1988.
- [43] Lei et al., J. Bacteriol., 169, 4379, 1987.
- [44] Verhoeyen et al., Science, 239, 1534, 1988.
- [45] Gronenbom, Mol. Gen. Genet, 148, 243, 1976.
- [46] Dagert et al., Gene, 6, 23, 1974.
- [47] Hanahan, J. Mol. Biol., 166, 557, 1983.
- [48] Jones et al., Nature, 321, 522, 1986.
- [49] Segal, Enzyme Kinetics, 73, Wiley, New York, 1975.
- [50] Gutfreund, Enzymes, Physical Principles, Wiley Interscience, London, 1972.
- [51] Chaidaroglou, Biochem., 27, 8338, 1988.
- [52] Grodberg and Dunn, J. Bacteriol., 170, 1245-1253, 1988.
Claims (32)
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GB888826444A GB8826444D0 (en) | 1988-11-11 | 1988-11-11 | Cloning immunoglobulin variable domains for expression by polymerase chain reaction |
GB8826444.5 | 1988-11-11 | ||
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US08/470,031 Expired - Lifetime US6248516B1 (en) | 1988-11-11 | 1995-06-06 | Single domain ligands, receptors comprising said ligands methods for their production, and use of said ligands and receptors |
US09/722,364 Expired - Fee Related US6545142B1 (en) | 1988-11-11 | 2000-11-28 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US10/290,252 Expired - Fee Related US7306907B2 (en) | 1988-11-11 | 2002-11-08 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US10/290,233 Abandoned US20040110941A2 (en) | 1988-11-11 | 2002-11-08 | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8940298B2 (en) | 2007-09-04 | 2015-01-27 | The Regents Of The University Of California | High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection |
US8940871B2 (en) | 2006-03-20 | 2015-01-27 | The Regents Of The University Of California | Engineered anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting |
Families Citing this family (1597)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5229272A (en) * | 1989-04-25 | 1993-07-20 | Igen, Inc. | Catalytic antibody components |
EP0368684B2 (en) * | 1988-11-11 | 2004-09-29 | Medical Research Council | Cloning immunoglobulin variable domain sequences. |
US5530101A (en) | 1988-12-28 | 1996-06-25 | Protein Design Labs, Inc. | Humanized immunoglobulins |
WO1990007713A1 (en) * | 1988-12-29 | 1990-07-12 | Cytogen Corporation | Molecular recognition units |
US5196510A (en) * | 1988-12-29 | 1993-03-23 | Cytogen Corporation | Molecular recognition units |
US5116964A (en) * | 1989-02-23 | 1992-05-26 | Genentech, Inc. | Hybrid immunoglobulins |
US6406697B1 (en) | 1989-02-23 | 2002-06-18 | Genentech, Inc. | Hybrid immunoglobulins |
US5225538A (en) * | 1989-02-23 | 1993-07-06 | Genentech, Inc. | Lymphocyte homing receptor/immunoglobulin fusion proteins |
DE3909799A1 (en) | 1989-03-24 | 1990-09-27 | Behringwerke Ag | MONOCLONAL ANTIBODIES (MAK) AGAINST TUMOR ASSOCIATED ANTIGENS, THEIR PRODUCTION AND USE |
US5236836A (en) * | 1989-04-25 | 1993-08-17 | Igen, Inc. | Autoantibodies which enhance the rate of a chemical reaction |
US5194585A (en) * | 1989-04-25 | 1993-03-16 | Igen, Inc. | Inhibitors of catalytic antibodies |
US5602015A (en) * | 1989-04-25 | 1997-02-11 | Igen, Inc. | Autoantibodies which enhance the rate of a chemical reaction |
US5318897A (en) * | 1989-04-25 | 1994-06-07 | Igen, Inc. | Monoclonal antibody and antibody components elicited to a polypeptide antigen ground state |
US5658753A (en) * | 1989-04-25 | 1997-08-19 | Paul; Sudhir | Catalytic antibody components |
US5599538A (en) * | 1989-04-25 | 1997-02-04 | Igen, Inc. | Autoantibodies which enhance the rate of a chemical reaction |
US6048717A (en) * | 1989-04-25 | 2000-04-11 | Igen International, Inc. | Inhibitors of catalytic antibodies |
CA2016841C (en) * | 1989-05-16 | 1999-09-21 | William D. Huse | A method for producing polymers having a preselected activity |
US6680192B1 (en) | 1989-05-16 | 2004-01-20 | Scripps Research Institute | Method for producing polymers having a preselected activity |
US6291161B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for tapping the immunological repertiore |
US6969586B1 (en) | 1989-05-16 | 2005-11-29 | Scripps Research Institute | Method for tapping the immunological repertoire |
US6291160B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for producing polymers having a preselected activity |
US6291158B1 (en) | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for tapping the immunological repertoire |
US6291159B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for producing polymers having a preselected activity |
US5231015A (en) * | 1989-10-18 | 1993-07-27 | Eastman Kodak Company | Methods of extracting nucleic acids and pcr amplification without using a proteolytic enzyme |
US6274324B1 (en) | 1989-12-01 | 2001-08-14 | Unilever Patent Holdings B.V. | Specific binding reagent comprising a variable domain protein linked to a support or tracer |
GB8928501D0 (en) * | 1989-12-18 | 1990-02-21 | Unilever Plc | Reagents |
ATE277179T1 (en) † | 1990-02-01 | 2004-10-15 | Dade Behring Marburg Gmbh | PRODUCTION AND USE OF HUMAN ANTIBODIES GENE BANKS (ßHUMAN ANTIBODIES LIBRARIESß) |
GB9021671D0 (en) * | 1990-10-05 | 1990-11-21 | Unilever Plc | Delivery of agents |
US5427908A (en) * | 1990-05-01 | 1995-06-27 | Affymax Technologies N.V. | Recombinant library screening methods |
US5723286A (en) | 1990-06-20 | 1998-03-03 | Affymax Technologies N.V. | Peptide library and screening systems |
US6172197B1 (en) | 1991-07-10 | 2001-01-09 | Medical Research Council | Methods for producing members of specific binding pairs |
US6916605B1 (en) | 1990-07-10 | 2005-07-12 | Medical Research Council | Methods for producing members of specific binding pairs |
GB9015198D0 (en) * | 1990-07-10 | 1990-08-29 | Brien Caroline J O | Binding substance |
GB9206318D0 (en) * | 1992-03-24 | 1992-05-06 | Cambridge Antibody Tech | Binding substances |
US7063943B1 (en) | 1990-07-10 | 2006-06-20 | Cambridge Antibody Technology | Methods for producing members of specific binding pairs |
GB9016299D0 (en) * | 1990-07-25 | 1990-09-12 | Brien Caroline J O | Binding substances |
DE4033120A1 (en) * | 1990-10-18 | 1992-04-23 | Boehringer Mannheim Gmbh | Genomic DNA fragment encoding antibody variable region prodn. - by attaching primers to hybridoma DNA then subjecting to polymerase chain reaction, for constructing genes encoding chimeric antibodies |
US5571894A (en) * | 1991-02-05 | 1996-11-05 | Ciba-Geigy Corporation | Recombinant antibodies specific for a growth factor receptor |
AU1025692A (en) * | 1991-02-06 | 1992-08-13 | Ciba-Geigy Ag | Novel chimeric antiidiotypic monoclonal antibodies |
CA2081028C (en) * | 1991-03-12 | 1999-12-14 | Barbara P. Wallner | Cd2 binding domain of lymphocyte function associated antigen 3 |
MX9203138A (en) * | 1991-03-12 | 1992-09-01 | Biogen Inc | DOMAIN OF LINK CD2-ANTIGEN 3 (LFA-3) ASSOCIATED WITH FUNCTION LYMPHOSITES. |
IE921169A1 (en) | 1991-04-10 | 1992-10-21 | Scripps Research Inst | Heterodimeric receptor libraries using phagemids |
US5962255A (en) * | 1992-03-24 | 1999-10-05 | Cambridge Antibody Technology Limited | Methods for producing recombinant vectors |
US6492160B1 (en) | 1991-05-15 | 2002-12-10 | Cambridge Antibody Technology Limited | Methods for producing members of specific binding pairs |
US6225447B1 (en) | 1991-05-15 | 2001-05-01 | Cambridge Antibody Technology Ltd. | Methods for producing members of specific binding pairs |
US5871907A (en) * | 1991-05-15 | 1999-02-16 | Medical Research Council | Methods for producing members of specific binding pairs |
US5858657A (en) * | 1992-05-15 | 1999-01-12 | Medical Research Council | Methods for producing members of specific binding pairs |
US6800738B1 (en) | 1991-06-14 | 2004-10-05 | Genentech, Inc. | Method for making humanized antibodies |
WO1994004679A1 (en) * | 1991-06-14 | 1994-03-03 | Genentech, Inc. | Method for making humanized antibodies |
LU91067I2 (en) | 1991-06-14 | 2004-04-02 | Genentech Inc | Trastuzumab and its variants and immunochemical derivatives including immotoxins |
US5939531A (en) * | 1991-07-15 | 1999-08-17 | Novartis Corp. | Recombinant antibodies specific for a growth factor receptor |
NL9101290A (en) * | 1991-07-23 | 1993-02-16 | Stichting Rega V Z W | RECOMBINANT DNA MOLECULA FOR THE EXPRESSION OF AN FV FRAGMENT OF AN ANTIBODY. |
US6764681B2 (en) | 1991-10-07 | 2004-07-20 | Biogen, Inc. | Method of prophylaxis or treatment of antigen presenting cell driven skin conditions using inhibitors of the CD2/LFA-3 interaction |
US5733731A (en) * | 1991-10-16 | 1998-03-31 | Affymax Technologies N.V. | Peptide library and screening method |
US5270170A (en) * | 1991-10-16 | 1993-12-14 | Affymax Technologies N.V. | Peptide library and screening method |
PT1024191E (en) | 1991-12-02 | 2008-12-22 | Medical Res Council | Production of anti-self antibodies from antibody segment repertoires and displayed on phage |
EP2224006A1 (en) * | 1991-12-02 | 2010-09-01 | MedImmune Limited | Production of anti-self antibodies from antibody segment repertoires and displayed on phage |
DE4142077A1 (en) * | 1991-12-19 | 1993-06-24 | Boehringer Mannheim Gmbh | METHOD FOR EXPRESSING RECOMBINANT ANTIKOERPERS |
US5824307A (en) | 1991-12-23 | 1998-10-20 | Medimmune, Inc. | Human-murine chimeric antibodies against respiratory syncytial virus |
US6399368B1 (en) | 1992-01-17 | 2002-06-04 | Board Of Regents, The University Of Texas System | Secretion of T cell receptor fragments from recombinant Escherichia coli cells |
SK280610B6 (en) | 1992-02-06 | 2000-05-16 | Schering Corporation | Humanised monoclonal antibodies against human interleukin-5 or a fragment thereof, hybridoma, polypeptide, isolated dna, recombinant vector, host cells, process for the preparation of polypeptide, and pharmaceutical compositions |
US5733743A (en) * | 1992-03-24 | 1998-03-31 | Cambridge Antibody Technology Limited | Methods for producing members of specific binding pairs |
CA2118508A1 (en) * | 1992-04-24 | 1993-11-11 | Elizabeth S. Ward | Recombinant production of immunoglobulin-like domains in prokaryotic cells |
GB9216983D0 (en) * | 1992-08-11 | 1992-09-23 | Unilever Plc | Polypeptide production |
US6765087B1 (en) * | 1992-08-21 | 2004-07-20 | Vrije Universiteit Brussel | Immunoglobulins devoid of light chains |
DE4233152A1 (en) * | 1992-10-02 | 1994-04-07 | Behringwerke Ag | Antibody-enzyme conjugates for prodrug activation |
GB9225453D0 (en) | 1992-12-04 | 1993-01-27 | Medical Res Council | Binding proteins |
ES2247204T3 (en) | 1994-01-31 | 2006-03-01 | Trustees Of Boston University | BANKS OF POLYCLONAL ANTIBODIES. |
US6010861A (en) * | 1994-08-03 | 2000-01-04 | Dgi Biotechnologies, Llc | Target specific screens and their use for discovering small organic molecular pharmacophores |
US6056957A (en) * | 1994-08-04 | 2000-05-02 | Schering Corporation | Humanized monoclonal antibodies against human interleukin-5 |
US6706484B1 (en) | 1995-08-18 | 2004-03-16 | Morphosys Ag | Protein/(poly)peptide libraries |
ES2176484T3 (en) * | 1995-08-18 | 2002-12-01 | Morphosys Ag | PROTEIN BANKS / (POLI) PEPTIDES. |
US7368111B2 (en) | 1995-10-06 | 2008-05-06 | Cambridge Antibody Technology Limited | Human antibodies specific for TGFβ2 |
US6136311A (en) | 1996-05-06 | 2000-10-24 | Cornell Research Foundation, Inc. | Treatment and diagnosis of cancer |
DK0898618T3 (en) * | 1996-05-10 | 2008-02-25 | Novozymes As | Method for providing novel DNA sequences |
PT1143006E (en) | 1996-08-19 | 2008-06-27 | Morphosys Ip Gmbh | Vectors/dna-sequences from human combinatorial antibody libraries |
GB9701425D0 (en) | 1997-01-24 | 1997-03-12 | Bioinvent Int Ab | A method for in vitro molecular evolution of protein function |
US6670453B2 (en) | 1997-10-27 | 2003-12-30 | Unilever Patent Holdings B.V. | Multivalent antigen-binding proteins |
EP1965213A3 (en) | 1998-02-04 | 2009-07-15 | Invitrogen Corporation | Microarrays and uses therefor |
EP1078051B1 (en) | 1998-05-13 | 2007-12-12 | Domantis Limited | Phage-display system for the selection of correctly folded proteins |
US6914128B1 (en) | 1999-03-25 | 2005-07-05 | Abbott Gmbh & Co. Kg | Human antibodies that bind human IL-12 and methods for producing |
US6492497B1 (en) * | 1999-04-30 | 2002-12-10 | Cambridge Antibody Technology Limited | Specific binding members for TGFbeta1 |
US7534605B2 (en) | 1999-06-08 | 2009-05-19 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | CD44 polypeptides, polynucleotides encoding same, antibodies directed thereagainst and method of using same for diagnosing and treating inflammatory diseases |
US7297478B1 (en) | 2000-09-22 | 2007-11-20 | Large Scale Biology Corporation | Creation of variable length and sequence linker regions for dual-domain or multi-domain molecules |
US6656467B2 (en) | 2000-01-27 | 2003-12-02 | Medimmune, Inc. | Ultra high affinity neutralizing antibodies |
AU2001234125A1 (en) | 2000-02-22 | 2001-09-03 | Medical And Biological Laboratories Co., Ltd. | Antibody library |
CA2401652A1 (en) | 2000-03-01 | 2001-09-07 | Medimmune, Inc. | High potency recombinant antibodies and method for producing them |
ES2393535T3 (en) * | 2000-04-17 | 2012-12-26 | Dyax Corp. | New methods to build libraries of genetic packages that collectively present members of a diverse family of peptides, polypeptides or proteins |
US8288322B2 (en) | 2000-04-17 | 2012-10-16 | Dyax Corp. | Methods of constructing libraries comprising displayed and/or expressed members of a diverse family of peptides, polypeptides or proteins and the novel libraries |
EP1176200A3 (en) | 2000-06-20 | 2005-01-12 | Switch Biotech Aktiengesellschaft | Use of polyeptides or their encoding nucleic acids for the diagnosis or treatment of skin diseases or wound healing and their use in indentifying pharmacologically acitve substances |
UY26807A1 (en) | 2000-06-29 | 2002-01-31 | Abbott Lab | DOUBLE SPECIFICITY ANTIBODIES AND METHODS FOR THE ELABORATION AND USE OF THE SAME |
US7288390B2 (en) | 2000-08-07 | 2007-10-30 | Centocor, Inc. | Anti-dual integrin antibodies, compositions, methods and uses |
US6902734B2 (en) | 2000-08-07 | 2005-06-07 | Centocor, Inc. | Anti-IL-12 antibodies and compositions thereof |
UA81743C2 (en) | 2000-08-07 | 2008-02-11 | Центокор, Инк. | HUMAN MONOCLONAL ANTIBODY WHICH SPECIFICALLY BINDS TUMOR NECROSIS FACTOR ALFA (TNFα), PHARMACEUTICAL MIXTURE CONTAINING THEREOF, AND METHOD FOR TREATING ARTHRITIS |
US20050196755A1 (en) * | 2000-11-17 | 2005-09-08 | Maurice Zauderer | In vitro methods of producing and identifying immunoglobulin molecules in eukaryotic cells |
CA2429544C (en) | 2000-11-17 | 2010-10-19 | University Of Rochester | In vitro methods of producing and identifying immunoglobulin molecules in eukaryotic cells |
US7179900B2 (en) | 2000-11-28 | 2007-02-20 | Medimmune, Inc. | Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment |
EP1366191A2 (en) | 2000-12-11 | 2003-12-03 | Alexion Pharmaceuticals, Inc. | Nested oligonucleotides containing hairpin for nucleic acid amplification |
PT1355919E (en) | 2000-12-12 | 2011-03-02 | Medimmune Llc | Molecules with extended half-lives, compositions and uses thereof |
US6958213B2 (en) | 2000-12-12 | 2005-10-25 | Alligator Bioscience Ab | Method for in vitro molecular evolution of protein function |
US7658921B2 (en) | 2000-12-12 | 2010-02-09 | Medimmune, Llc | Molecules with extended half-lives, compositions and uses thereof |
AU2002249854B2 (en) | 2000-12-18 | 2007-09-20 | Dyax Corp. | Focused libraries of genetic packages |
JP4986370B2 (en) | 2000-12-22 | 2012-07-25 | マックス−プランク−ゲゼルシャフト・ツア・フェルデルング・デア・ヴィッセンシャフテン・エー・ファオ | Uses of RGM and its modulators |
US20020086292A1 (en) | 2000-12-22 | 2002-07-04 | Shigeaki Harayama | Synthesis of hybrid polynucleotide molecules using single-stranded polynucleotide molecules |
AU2002338446A1 (en) * | 2001-01-23 | 2002-11-05 | University Of Rochester Medical Center | Methods of producing or identifying intrabodies in eukaryotic cells |
EP2067486A1 (en) | 2001-01-31 | 2009-06-10 | Biogen Idec Inc. | Use of CD23 antagonists for the treatment of neoplastic disorders |
GB0110029D0 (en) | 2001-04-24 | 2001-06-13 | Grosveld Frank | Transgenic animal |
US6972324B2 (en) | 2001-05-18 | 2005-12-06 | Boehringer Ingelheim Pharmaceuticals, Inc. | Antibodies specific for CD44v6 |
WO2004058821A2 (en) * | 2002-12-27 | 2004-07-15 | Domantis Limited | Dual specific single domain antibodies specific for a ligand and for the receptor of the ligand |
GB0115841D0 (en) | 2001-06-28 | 2001-08-22 | Medical Res Council | Ligand |
WO2004003019A2 (en) * | 2002-06-28 | 2004-01-08 | Domantis Limited | Immunoglobin single variant antigen-binding domains and dual-specific constructs |
EP1399484B1 (en) * | 2001-06-28 | 2010-08-11 | Domantis Limited | Dual-specific ligand and its use |
US20060073141A1 (en) * | 2001-06-28 | 2006-04-06 | Domantis Limited | Compositions and methods for treating inflammatory disorders |
AU2002320352A1 (en) | 2001-07-24 | 2003-02-17 | Biogen Idec Ma Inc. | Methods for treating or preventing sclerotic disorders using cd2-binding agents |
US6833441B2 (en) | 2001-08-01 | 2004-12-21 | Abmaxis, Inc. | Compositions and methods for generating chimeric heteromultimers |
EP2277914A3 (en) | 2001-08-10 | 2012-08-08 | Aberdeen University | Antigen binding domains from fish |
WO2003018749A2 (en) * | 2001-08-22 | 2003-03-06 | Shengfeng Li | Compositions and methods for generating antigen-binding units |
US20040005709A1 (en) * | 2001-10-24 | 2004-01-08 | Hoogenboom Henricus Renerus Jacobus Mattheus | Hybridization control of sequence variation |
US7175983B2 (en) | 2001-11-02 | 2007-02-13 | Abmaxis, Inc. | Adapter-directed display systems |
GB0126378D0 (en) | 2001-11-02 | 2002-01-02 | Oxford Biomedica Ltd | Antigen |
EP1456237A2 (en) * | 2001-12-21 | 2004-09-15 | Vlaams Interuniversitair Instituut voor Biotechnologie vzw. | Method for cloning of variable domain sequences |
EP2075256A2 (en) | 2002-01-14 | 2009-07-01 | William Herman | Multispecific binding molecules |
SI3031910T1 (en) | 2002-02-21 | 2018-05-31 | Institute Of Virology Slovak Academy Of Sciences | Mn/ca ix-specific monoclonal antibodies generated from mn/ca ix-deficient mice and methods of use |
US7718776B2 (en) * | 2002-04-05 | 2010-05-18 | Amgen Inc. | Human anti-OPGL neutralizing antibodies as selective OPGL pathway inhibitors |
US7135310B2 (en) | 2002-04-24 | 2006-11-14 | The Regents Of The University Of California | Method to amplify variable sequences without imposing primer sequences |
EP2314622B1 (en) | 2002-05-22 | 2017-10-18 | ESBATech, an Alcon Biomedical Research Unit LLC | Immunoglobulin frameworks which demonstrate enhanced stability in the intracellular environment and methods of identifying same |
NZ556507A (en) | 2002-06-03 | 2010-03-26 | Genentech Inc | Synthetic antibody phage libraries |
EP2305710A3 (en) | 2002-06-03 | 2013-05-29 | Genentech, Inc. | Synthetic antibody phage libraries |
US7132100B2 (en) | 2002-06-14 | 2006-11-07 | Medimmune, Inc. | Stabilized liquid anti-RSV antibody formulations |
GB0213745D0 (en) | 2002-06-14 | 2002-07-24 | Univ Edinburgh | Enzyme |
US7425618B2 (en) | 2002-06-14 | 2008-09-16 | Medimmune, Inc. | Stabilized anti-respiratory syncytial virus (RSV) antibody formulations |
US9028822B2 (en) | 2002-06-28 | 2015-05-12 | Domantis Limited | Antagonists against TNFR1 and methods of use therefor |
US9321832B2 (en) | 2002-06-28 | 2016-04-26 | Domantis Limited | Ligand |
US7696320B2 (en) | 2004-08-24 | 2010-04-13 | Domantis Limited | Ligands that have binding specificity for VEGF and/or EGFR and methods of use therefor |
US20060002935A1 (en) | 2002-06-28 | 2006-01-05 | Domantis Limited | Tumor Necrosis Factor Receptor 1 antagonists and methods of use therefor |
EP1527346B1 (en) | 2002-08-07 | 2011-06-08 | Ablynx N.V. | Modulation of platelet adhesion based on the surface exposed beta-switch loop of platelet glycoprotein ib-alpha |
US20040067532A1 (en) | 2002-08-12 | 2004-04-08 | Genetastix Corporation | High throughput generation and affinity maturation of humanized antibody |
MEP32508A (en) * | 2002-09-06 | 2010-10-10 | Amgen Inc | Therapeutic human anti-il-1r1 monoclonal antibody |
ATE425183T1 (en) | 2002-10-07 | 2009-03-15 | Ludwig Inst For Cancer Res Ltd | P53 BINDING POLYPEPTIDE |
WO2004034988A2 (en) | 2002-10-16 | 2004-04-29 | Amgen Inc. | Human anti-ifn-ϝ neutralizing antibodies as selective ifn-ϝ pathway inhibitors |
US9701754B1 (en) | 2002-10-23 | 2017-07-11 | City Of Hope | Covalent disulfide-linked diabodies and uses thereof |
JP2006520584A (en) | 2002-11-08 | 2006-09-14 | アブリンクス エン.ヴェー. | Stabilized single domain antibody |
US9320792B2 (en) | 2002-11-08 | 2016-04-26 | Ablynx N.V. | Pulmonary administration of immunoglobulin single variable domains and constructs thereof |
US20060034845A1 (en) | 2002-11-08 | 2006-02-16 | Karen Silence | Single domain antibodies directed against tumor necrosis factor alpha and uses therefor |
GB0230203D0 (en) * | 2002-12-27 | 2003-02-05 | Domantis Ltd | Fc fusion |
GB0230201D0 (en) * | 2002-12-27 | 2003-02-05 | Domantis Ltd | Retargeting |
MXPA05006043A (en) | 2003-01-10 | 2006-01-30 | Ablynx Nv | Therapeutic polypeptides, homologues thereof, fragments thereof and for use in modulating platelet-mediated aggregation. |
AU2004207741B2 (en) * | 2003-01-24 | 2011-02-10 | Applied Molecular Evolution, Inc | Human IL-1 beta antagonists |
DE10303974A1 (en) | 2003-01-31 | 2004-08-05 | Abbott Gmbh & Co. Kg | Amyloid β (1-42) oligomers, process for their preparation and their use |
PT2248899E (en) | 2003-03-19 | 2015-09-23 | Biogen Ma Inc | Nogo receptor binding protein |
TWI353991B (en) | 2003-05-06 | 2011-12-11 | Syntonix Pharmaceuticals Inc | Immunoglobulin chimeric monomer-dimer hybrids |
US9708410B2 (en) | 2003-05-30 | 2017-07-18 | Janssen Biotech, Inc. | Anti-tissue factor antibodies and compositions |
EP1498133A1 (en) | 2003-07-18 | 2005-01-19 | Aventis Pharma Deutschland GmbH | Use of a pak inhibitor for the treatment of a joint disease |
US20050106667A1 (en) | 2003-08-01 | 2005-05-19 | Genentech, Inc | Binding polypeptides with restricted diversity sequences |
CA2534055A1 (en) * | 2003-08-01 | 2005-02-10 | Genentech, Inc. | Antibody cdr polypeptide sequences with restricted diversity |
US7758859B2 (en) | 2003-08-01 | 2010-07-20 | Genentech, Inc. | Anti-VEGF antibodies |
US20120202710A1 (en) * | 2003-09-09 | 2012-08-09 | Integrigen, Inc. | Methods and compositions for generation of germline human antibody genes |
ES2339710T5 (en) | 2003-09-23 | 2017-10-05 | University Of North Carolina At Chapel Hill | Cells that coexpress vitamin K reductase and vitamin K dependent protein and use them to improve the productivity of said vitamin K dependent protein |
EP2272951B1 (en) | 2003-10-14 | 2014-07-23 | Baxter International Inc. | Vitamin k epoxide recycling polypeptide vkorc1, a therapeutic target of coumarin and their derivatives |
EP1697415A1 (en) | 2003-11-12 | 2006-09-06 | Biogen Idec MA Inc. | NEONATAL Fc RECEPTOR (FcRn)-BINDING POLYPEPTIDE VARIANTS, DIMERIC Fc BINDING PROTEINS AND METHODS RELATED THERETO |
CA2544577C (en) | 2003-12-01 | 2013-01-08 | Dako Denmark A/S | Methods and compositions for immuno-histochemical detection |
GB0328690D0 (en) | 2003-12-10 | 2004-01-14 | Ludwig Inst Cancer Res | Tumour suppressor protein |
DK1711528T3 (en) | 2003-12-23 | 2012-08-20 | Genentech Inc | TREATMENT OF CANCER WITH UNTIL UNKNOWN ANTI-IL 13 MONOCLONAL ANTIBODIES |
AU2005207003C1 (en) * | 2004-01-20 | 2013-06-13 | Humanigen, Inc. | Antibody specificity transfer using minimal essential binding determinants |
GB0406215D0 (en) | 2004-03-19 | 2004-04-21 | Procure Therapeutics Ltd | Prostate stem cell |
RU2429245C2 (en) | 2004-03-30 | 2011-09-20 | Глаксо Груп Лимитед | Immunoglobulins |
US7785903B2 (en) | 2004-04-09 | 2010-08-31 | Genentech, Inc. | Variable domain library and uses |
ES2442386T3 (en) | 2004-04-23 | 2014-02-11 | Bundesrepublik Deutschland Letztvertreten Durch Das Robert Koch-Institut Vertreten Durch Seinen Pr | Method for the treatment of conditions mediated by T cells by the decrease of positive ICOS cells in vivo. |
US7662921B2 (en) | 2004-05-07 | 2010-02-16 | Astellas Us Llc | Methods of treating viral disorders |
EA012622B1 (en) * | 2004-06-01 | 2009-10-30 | Домэнтис Лимитед | Bispecific fusion antibodies with enhanced serum half-life |
US7977071B2 (en) | 2004-06-02 | 2011-07-12 | Adalta Pty Ltd. | Binding moieties based on shark ignar domains |
PT1776136E (en) | 2004-06-24 | 2012-12-05 | Biogen Idec Inc | Treatment of conditions involving demyelination |
TWI307630B (en) | 2004-07-01 | 2009-03-21 | Glaxo Group Ltd | Immunoglobulins |
GB0414886D0 (en) | 2004-07-02 | 2004-08-04 | Neutec Pharma Plc | Treatment of bacterial infections |
AU2005263334C1 (en) | 2004-07-20 | 2011-01-20 | Symphogen A/S | A procedure for structural characterization of a recombinant polyclonal protein or a polyclonal cell line |
MX2007000644A (en) | 2004-07-20 | 2007-03-28 | Symphogen As | Anti-rhesus d recombinant polyclonal antibody and methods of manufacture. |
EP1864998B2 (en) | 2004-07-22 | 2022-06-22 | Erasmus University Medical Center Rotterdam | Binding molecules |
GB0416487D0 (en) | 2004-07-23 | 2004-08-25 | Isis Innovation | Modified virus |
US7846438B2 (en) | 2004-08-03 | 2010-12-07 | Biogen Idec Ma Inc. | Methods of promoting neurite outgrowth with soluble TAJ polypeptides |
US7563443B2 (en) | 2004-09-17 | 2009-07-21 | Domantis Limited | Monovalent anti-CD40L antibody polypeptides and compositions thereof |
PL3540062T3 (en) * | 2004-11-16 | 2021-12-27 | Humanigen, Inc. | Immunoglobulin variable region cassette exchange |
GB0425739D0 (en) * | 2004-11-23 | 2004-12-22 | Procure Therapeutics Ltd | Humanised baculovirus 2 |
GB0521621D0 (en) | 2005-10-24 | 2005-11-30 | Domantis Ltd | Tumor necrosis factor receptor 1 antagonists for treating respiratory diseases |
FR2879605B1 (en) | 2004-12-16 | 2008-10-17 | Centre Nat Rech Scient Cnrse | PRODUCTION OF ANTIBODY FORMATS AND IMMUNOLOGICAL APPLICATIONS OF THESE FORMATS |
WO2006071200A2 (en) | 2004-12-30 | 2006-07-06 | Agency For Science, Technology And Research | Chinese hamster apoptosis-related genes |
EP2311880A3 (en) | 2005-01-05 | 2011-07-27 | Biogen Idec MA Inc. | Cripto binding molecules |
ATE473446T1 (en) | 2005-01-14 | 2010-07-15 | Ablynx Nv | METHODS AND TEST DEVICES FOR DIFFERENTIATING DIFFERENT FORMS OF DISEASES AND DISEASES CHARACTERIZED BY THROMBOCYTOPENIA AND/OR BY SPONTANEOUS INTERACTIONS BETWEEN THE VON WILLEBRAND FACTOR AND PLATES |
AU2006222204B2 (en) | 2005-03-11 | 2012-09-27 | Sanofi-Aventis | Use of MGC4504 |
CA2601574C (en) | 2005-03-15 | 2014-12-02 | University Of North Carolina At Chapel Hill | Methods and compositions for producing active vitamin k-dependent proteins |
JP2008532559A (en) | 2005-03-19 | 2008-08-21 | メディカル リサーチ カウンシル | Treatment and prevention of viral infection or improvement of treatment and prevention |
PT2343320T (en) | 2005-03-25 | 2018-01-23 | Gitr Inc | Anti-gitr antibodies and uses thereof |
EP1882044A1 (en) | 2005-05-11 | 2008-01-30 | sanofi-aventis | Use of a gip promoter polymorphism |
PL1888641T3 (en) | 2005-05-18 | 2012-05-31 | Ablynx Nv | Serum albumin binding proteins |
PE20061444A1 (en) | 2005-05-19 | 2007-01-15 | Centocor Inc | ANTI-MCP-1 ANTIBODY, COMPOSITIONS, METHODS AND USES |
HUE039846T2 (en) | 2005-05-20 | 2019-02-28 | Ablynx Nv | Improved nanobodies tm for the treatment of aggregation-mediated disorders |
WO2006138739A2 (en) | 2005-06-17 | 2006-12-28 | Tolerx, Inc. | Ilt3 binding molecules and uses therefor |
CN103172739A (en) | 2005-06-30 | 2013-06-26 | Abbvie公司 | IL-12/P40 binding proteins |
SI1904104T1 (en) | 2005-07-08 | 2013-12-31 | Biogen Idec Ma Inc. | Sp35 antibodies and uses thereof |
KR20140053410A (en) | 2005-08-19 | 2014-05-07 | 아보트 러보러터리즈 | Dual variable domain immunoglobulin and uses thereof |
US7612181B2 (en) | 2005-08-19 | 2009-11-03 | Abbott Laboratories | Dual variable domain immunoglobulin and uses thereof |
EP2500357A3 (en) | 2005-08-19 | 2012-10-24 | Abbott Laboratories | Dual variable domain immunoglobulin and uses thereof |
RS55788B1 (en) | 2005-08-31 | 2017-08-31 | Merck Sharp & Dohme | Engineered anti-il-23 antibodies |
JP2009510002A (en) | 2005-09-30 | 2009-03-12 | アボット ゲゼルシャフト ミット ベシュレンクテル ハフツング ウント コンパニー コマンディトゲゼルシャフト | Binding domains of proteins of the repulsion-inducing molecule (RGM) protein family, and functional fragments thereof, and uses thereof |
JP5270352B2 (en) | 2005-10-03 | 2013-08-21 | スミス アンド ネフュー インコーポレーテッド | Fixture assembly |
DE102005048898A1 (en) | 2005-10-12 | 2007-04-19 | Sanofi-Aventis Deutschland Gmbh | EGLN2 variants and their use in the prevention or treatment of thromboembolic disorders and coronary heart disease |
GB0521139D0 (en) | 2005-10-18 | 2005-11-23 | Univ Sheffield | Therapeutic agent |
MX2008005405A (en) * | 2005-10-28 | 2008-09-11 | Florida Internat University Bo | Horse: human chimeric antibodies. |
EP1959979A4 (en) | 2005-11-04 | 2010-01-27 | Biogen Idec Inc | Methods for promoting neurite outgrowth and survival of dopaminergic neurons |
EP3299027A1 (en) | 2005-11-04 | 2018-03-28 | Genentech, Inc. | Use of complement pathway inhibitors to treat ocular diseases |
EP2465870A1 (en) | 2005-11-07 | 2012-06-20 | Genentech, Inc. | Binding polypeptides with diversified and consensus VH/VL hypervariable sequences |
UA96139C2 (en) | 2005-11-08 | 2011-10-10 | Дженентек, Інк. | Anti-neuropilin-1 (nrp1) antibody |
AU2006318580A1 (en) | 2005-11-21 | 2007-05-31 | Merck Serono Sa | Compositions and methods of producing hybrid antigen binding molecules and uses thereof |
JP5475994B2 (en) | 2005-11-30 | 2014-04-16 | アッヴィ・インコーポレイテッド | Anti-Aβ globulomer antibody, antigen-binding portion thereof, corresponding hybridoma, nucleic acid, vector, host cell, method for producing said antibody, composition comprising said antibody, use of said antibody and method of using said antibody. |
RS53270B2 (en) | 2005-11-30 | 2018-05-31 | Abbvie Deutschland | Monoclonal antibodies against amyloid beta protein and uses thereof |
EP1957536A2 (en) * | 2005-12-01 | 2008-08-20 | Domantis Limited | Noncompetitive domain antibody formats that bind interleukin 1 receptor type 1 |
EP1973951A2 (en) * | 2005-12-02 | 2008-10-01 | Genentech, Inc. | Binding polypeptides with restricted diversity sequences |
RU2470941C2 (en) | 2005-12-02 | 2012-12-27 | Дженентек, Инк. | Binding polypeptides and use thereof |
US7737259B2 (en) | 2005-12-02 | 2010-06-15 | Genentech, Inc. | Compositions and methods for the treatment of diseases and disorders associated with cytokine signaling |
EP1965827B1 (en) | 2005-12-02 | 2015-02-25 | Biogen Idec MA Inc. | Treatment of conditions involving demyelination |
ES2550099T3 (en) | 2006-01-27 | 2015-11-04 | Biogen Ma Inc. | Nogo receptor antagonists |
EP1987064A4 (en) | 2006-02-01 | 2010-04-07 | Arana Therapeutics Ltd | Domain antibody construct |
EP2010677A4 (en) | 2006-02-24 | 2010-04-14 | Investigen Inc | Methods and compositions for detecting polynucleotides |
WO2007110219A1 (en) * | 2006-03-27 | 2007-10-04 | Ablynx N.V. | Medical delivery device for therapeutic proteins based on single domain antibodies |
EP2016101A2 (en) * | 2006-05-09 | 2009-01-21 | Genentech, Inc. | Binding polypeptides with optimized scaffolds |
US8524865B2 (en) | 2006-05-30 | 2013-09-03 | Genentech, Inc. | Antibodies and immunoconjugates and uses therefor |
GB0611116D0 (en) | 2006-06-06 | 2006-07-19 | Oxford Genome Sciences Uk Ltd | Proteins |
WO2010123874A1 (en) | 2009-04-20 | 2010-10-28 | Oxford Biotherapeutics Ltd. | Antibodies specific to cadherin-17 |
US7777008B2 (en) | 2006-06-19 | 2010-08-17 | Tolerx, Inc. | ILT3 binding molecules and uses therefor |
US8874380B2 (en) | 2010-12-09 | 2014-10-28 | Rutgers, The State University Of New Jersey | Method of overcoming therapeutic limitations of nonuniform distribution of radiopharmaceuticals and chemotherapy drugs |
CN101563365B (en) * | 2006-08-03 | 2012-10-31 | 瓦西尼斯公司 | Anti-IL-6 monoclonal antibodies and uses thereof |
DK2059533T3 (en) | 2006-08-30 | 2013-02-25 | Genentech Inc | MULTI-SPECIFIC ANTIBODIES |
MX2009002418A (en) | 2006-09-05 | 2009-04-23 | Medarex Inc | Antibodies to bone morphogenic proteins and receptors therefor and methods for their use. |
SG174782A1 (en) | 2006-09-08 | 2011-10-28 | Abbott Lab | Interleukin - 13 binding proteins |
US20100008910A1 (en) | 2006-09-12 | 2010-01-14 | John Chant | Methods and compositions for the diagnosis and treatment of cancer |
PT2486941T (en) | 2006-10-02 | 2017-05-30 | Squibb & Sons Llc | Human antibodies that bind cxcr4 and uses thereof |
US7767206B2 (en) | 2006-10-02 | 2010-08-03 | Amgen Inc. | Neutralizing determinants of IL-17 Receptor A and antibodies that bind thereto |
GB0620705D0 (en) | 2006-10-18 | 2006-11-29 | Opsona Therapeutics | Compounds for the modulation of toll-like receptor activity and assay methods for the identification of said compounds |
KR101541550B1 (en) | 2006-10-27 | 2015-08-04 | 제넨테크, 인크. | Antibodies and immunoconjugates and uses therefor |
US8455626B2 (en) | 2006-11-30 | 2013-06-04 | Abbott Laboratories | Aβ conformer selective anti-aβ globulomer monoclonal antibodies |
US8067179B2 (en) | 2006-11-30 | 2011-11-29 | Research Development Foundation | Immunoglobulin libraries |
CN103172743B (en) | 2006-12-01 | 2015-04-08 | 梅达雷克斯有限责任公司 | Human Antibodies That Bind Cd22 And Uses Thereof |
CL2007003622A1 (en) | 2006-12-13 | 2009-08-07 | Medarex Inc | Human anti-cd19 monoclonal antibody; composition comprising it; and tumor cell growth inhibition method. |
KR20090088950A (en) | 2006-12-14 | 2009-08-20 | 쉐링 코포레이션 | Engineered anti-tslp antibody |
AU2007333098A1 (en) | 2006-12-14 | 2008-06-19 | Medarex, Inc. | Human antibodies that bind CD70 and uses thereof |
TWI419904B (en) | 2006-12-18 | 2013-12-21 | Genentech Inc | Antagonist anti-notch3 antibodies and their use in the prevention and treatment of notch3-related diseases |
EP2102244A2 (en) | 2006-12-19 | 2009-09-23 | Ablynx N.V. | Amino acid sequences directed against a metalloproteinase from the adam family and polypeptides comprising the same for the treatment of adam-related diseases and disorders |
US9512236B2 (en) | 2006-12-19 | 2016-12-06 | Ablynx N.V. | Amino acid sequences directed against GPCRS and polypeptides comprising the same for the treatment of GPCR-related diseases and disorders |
CA2673331A1 (en) | 2006-12-19 | 2008-06-26 | Ablynx N.V. | Amino acid sequences directed against gpcrs and polypeptides comprising the same for the treatment of gpcr-related diseases and disorders |
US8128926B2 (en) | 2007-01-09 | 2012-03-06 | Biogen Idec Ma Inc. | Sp35 antibodies and uses thereof |
NZ577976A (en) | 2007-01-09 | 2011-12-22 | Biogen Idec Inc | Sp35 antibodies and uses thereof |
WO2008088823A2 (en) | 2007-01-16 | 2008-07-24 | Abbott Laboratories | Methods for treating psoriasis |
US8664364B2 (en) | 2007-01-24 | 2014-03-04 | Carnegie Mellon University | Optical biosensors |
WO2008103702A2 (en) | 2007-02-23 | 2008-08-28 | Investigen, Inc. | Methods and compositions for rapid light-activated isolation and detection of analytes |
RS52345B (en) | 2007-02-23 | 2012-12-31 | Schering Corporation | Engineered anti-il-23p19 antibodies |
WO2008103473A1 (en) | 2007-02-23 | 2008-08-28 | Schering Corporation | Engineered anti-il-23p19 antibodies |
EP2121745A2 (en) | 2007-02-26 | 2009-11-25 | Oxford Genome Sciences (UK) Limited | Proteins |
WO2008104803A2 (en) | 2007-02-26 | 2008-09-04 | Oxford Genome Sciences (Uk) Limited | Proteins |
EP2124952A2 (en) | 2007-02-27 | 2009-12-02 | Abbott GmbH & Co. KG | Method for the treatment of amyloidoses |
CN101675076B (en) | 2007-02-28 | 2013-09-18 | 默沙东公司 | Engineered anti-il-23r antibodies |
CA2681581A1 (en) | 2007-03-30 | 2008-10-09 | Abbott Laboratories | Recombinant expression vector elements (reves) for enhancing expression of recombinant proteins in host cells |
US9969797B2 (en) * | 2008-04-23 | 2018-05-15 | Covalent Bioscience Incorporated | Immunoglobulins directed to bacterial, viral and endogenous polypeptides |
WO2008132516A1 (en) | 2007-04-26 | 2008-11-06 | Opsona Therapeutics Limited | Toll-like receptor binding epitope and compositions for binding thereto |
TWI570135B (en) | 2007-04-27 | 2017-02-11 | 建南德克公司 | Potent, stable and non-immunosuppressive anti-cd4 antibodies |
WO2008137475A2 (en) * | 2007-05-01 | 2008-11-13 | Research Development Foundation | Immunoglobulin fc libraries |
KR101523788B1 (en) | 2007-05-17 | 2015-06-26 | 제넨테크, 인크. | Crystal structures of neuropilin fragments and neuropilin-antibody complexes |
EP1997830A1 (en) | 2007-06-01 | 2008-12-03 | AIMM Therapeutics B.V. | RSV specific binding molecules and means for producing them |
NZ580530A (en) * | 2007-06-06 | 2012-04-27 | Domantis Ltd | Anti vegf polypeptides, antibody variable domains and antagonists |
CA2688434A1 (en) | 2007-06-06 | 2008-12-11 | Domantis Limited | Polypeptides, antibody variable domains and antagonists |
GB0724331D0 (en) | 2007-12-13 | 2008-01-23 | Domantis Ltd | Compositions for pulmonary delivery |
US8138313B2 (en) | 2007-06-15 | 2012-03-20 | Deutsches Krebsforschungszentrum Stiftung Des Offentlichen Rechts | Treatment of tumors using specific anti-L1 antibody |
US8263072B2 (en) | 2007-06-22 | 2012-09-11 | Genera Istrazivanja, d.o.o. | ADAMTS4 as a blood biomarker and therapeutic target for chronic renal failure |
EP2173772A2 (en) | 2007-07-03 | 2010-04-14 | Ablynx N.V. | Providing improved immunoglobulin sequences by mutating cdr and/or fr positions |
CN101801413A (en) | 2007-07-12 | 2010-08-11 | 托勒克斯股份有限公司 | Combination therapies employing GITR binding molecules |
EP2023144A1 (en) | 2007-08-01 | 2009-02-11 | Sanofi-Aventis | Novel AS160-like protein, test systems, methods and uses involving it for the identification of diabetes type 2 therapeutics |
US8734788B2 (en) | 2007-08-03 | 2014-05-27 | Opsona Therapeutics Ltd | Composition and method for treatment of reperfusion injury and tissue damage |
RU2015147300A (en) | 2007-08-29 | 2019-01-11 | Санофи-Авентис | HUMANIZED ANTIBODIES TO CXCR5, THEIR DERIVATIVES AND THEIR APPLICATION |
DK3124497T3 (en) | 2007-09-14 | 2020-05-11 | Adimab Llc | RATIONALE DESIGNED SYNTHETIC ANTIBODY LIBRARIES AND APPLICATIONS THEREOF |
US8877688B2 (en) * | 2007-09-14 | 2014-11-04 | Adimab, Llc | Rationally designed, synthetic antibody libraries and uses therefor |
ES2624450T3 (en) | 2007-09-18 | 2017-07-14 | Dako Denmark A/S | Fast and sensitive method for the detection of biological targets |
CA2700714C (en) | 2007-09-26 | 2018-09-11 | Ucb Pharma S.A. | Dual specificity antibody fusions |
PL3059246T3 (en) | 2007-09-26 | 2018-11-30 | Chugai Seiyaku Kabushiki Kaisha | Modified antibody constant region |
EP2050764A1 (en) | 2007-10-15 | 2009-04-22 | sanofi-aventis | Novel polyvalent bispecific antibody format and uses thereof |
JP5985150B2 (en) | 2007-11-07 | 2016-09-06 | ジェネンテック, インコーポレイテッド | Compositions and methods for the treatment of microbial disorders |
EP3115469B1 (en) | 2007-11-19 | 2020-04-29 | Celera Corporation | Lung cancer markers and uses thereof |
MX2010005324A (en) | 2007-11-19 | 2010-06-01 | Genentech Inc | Compositions and methods for inhibiting tumor progression. |
AU2008329466B2 (en) | 2007-11-27 | 2015-04-09 | The University Of British Columbia | 14-3-3 antagonists for the prevention and treatment of arthritis |
EP2215123A1 (en) | 2007-11-27 | 2010-08-11 | Ablynx N.V. | Immunoglobulin constructs |
TWI468417B (en) | 2007-11-30 | 2015-01-11 | Genentech Inc | Anti-vegf antibodies |
US8426153B2 (en) | 2007-12-03 | 2013-04-23 | Carnegie Mellon University | Linked peptides fluorogenic biosensors |
EP2594590B1 (en) | 2007-12-14 | 2014-11-12 | Bristol-Myers Squibb Company | Method of producing binding molecules for the human OX40 receptor |
US8574577B2 (en) | 2008-01-03 | 2013-11-05 | The Scripps Research Institute | VEGF antibodies comprising modular recognition domains |
US8454960B2 (en) | 2008-01-03 | 2013-06-04 | The Scripps Research Institute | Multispecific antibody targeting and multivalency through modular recognition domains |
NZ586701A (en) | 2008-01-03 | 2013-07-26 | Scripps Research Inst | Antibody targeting through a modular recognition domain (MRD) wherein the MRD targets angiopoietin-2 (ANG-2) |
US8557243B2 (en) | 2008-01-03 | 2013-10-15 | The Scripps Research Institute | EFGR antibodies comprising modular recognition domains |
US8557242B2 (en) | 2008-01-03 | 2013-10-15 | The Scripps Research Institute | ERBB2 antibodies comprising modular recognition domains |
US8962803B2 (en) | 2008-02-29 | 2015-02-24 | AbbVie Deutschland GmbH & Co. KG | Antibodies against the RGM A protein and uses thereof |
EP2098536A1 (en) | 2008-03-05 | 2009-09-09 | 4-Antibody AG | Isolation and identification of antigen- or ligand-specific binding proteins |
EP2247616A2 (en) | 2008-03-05 | 2010-11-10 | Ablynx N.V. | Novel antigen binding dimer-complexes, methods of making and uses thereof |
US9873957B2 (en) | 2008-03-13 | 2018-01-23 | Dyax Corp. | Libraries of genetic packages comprising novel HC CDR3 designs |
TW201513883A (en) | 2008-03-18 | 2015-04-16 | Abbvie Inc | Methods for treating psoriasis |
EP2105742A1 (en) | 2008-03-26 | 2009-09-30 | Sanofi-Aventis | Use of cathepsin C |
AU2009228158B2 (en) | 2008-03-27 | 2014-02-27 | Zymogenetics, Inc. | Compositions and methods for inhibiting PDGFRbeta and VEGF-A |
MX348362B (en) * | 2008-03-31 | 2017-06-07 | Genentech Inc * | Compositions and methods for treating and diagnosing asthma. |
AU2009235467A1 (en) | 2008-04-07 | 2009-10-15 | Ablynx Nv | Single variable domains against the Notch pathways |
AU2008201871A1 (en) * | 2008-04-16 | 2009-11-26 | Deutsches Krebsforschungszentrum Stiftung Des Oeffentlichen Rechts | Inhibition of angiogenesis and tumor metastasis |
JP5780951B2 (en) | 2008-04-24 | 2015-09-16 | ダイアックス コーポレーション | A library of genetic packages containing new HCCR1, CDR2 and CDR3 designs, and new LCCR1, CDR2 and CDR3 designs |
US9029508B2 (en) | 2008-04-29 | 2015-05-12 | Abbvie Inc. | Dual variable domain immunoglobulins and uses thereof |
CN102089430B (en) | 2008-05-09 | 2015-02-04 | Abbvie公司 | Antibodies to receptor of advanced glycation end products (RAGE) and uses thereof |
SG176464A1 (en) | 2008-05-09 | 2011-12-29 | Agency Science Tech & Res | Diagnosis and treatment of kawasaki disease |
BRPI0911984A2 (en) | 2008-05-16 | 2016-09-20 | Ablynx Nv | amino acid sequences directed against cxcr4 and other gpcrs compounds comprising the same |
CN102112494A (en) | 2008-06-03 | 2011-06-29 | 雅培制药有限公司 | Dual variable domain immunoglobulins and uses thereof |
EP3002299A1 (en) | 2008-06-03 | 2016-04-06 | AbbVie Inc. | Dual variable domain immunoglobulins and uses thereof |
DK2285408T3 (en) | 2008-06-05 | 2019-02-04 | Ablynx Nv | AMINO ACID SEQUENCES AGAINST COATING PROTEINS IN A VIRUS AND POLYPEPTIDES INCLUDING THESE FOR TREATMENT OF VIRUSAL DISEASES |
EP2650014A2 (en) | 2008-06-20 | 2013-10-16 | Wyeth LLC | Compositions and methods of use of ORF1358 from beta-hemolytic streptococcal strains |
EP2310049A4 (en) | 2008-07-08 | 2013-06-26 | Abbvie Inc | Prostaglandin e2 binding proteins and uses thereof |
KR20110031369A (en) | 2008-07-08 | 2011-03-25 | 아보트 러보러터리즈 | Prostaglandin e2 dual variable domain immunoglobulins and uses thereof |
EP2315779A2 (en) | 2008-07-09 | 2011-05-04 | Biogen Idec MA Inc. | Compositions comprising antibodies to lingo or fragments thereof |
EP2316030B1 (en) | 2008-07-25 | 2019-08-21 | Wagner, Richard W. | Protein screeing methods |
US9580513B2 (en) | 2008-08-08 | 2017-02-28 | Agency For Science, Technology And Research (A*Star) | VHZ for diagnosis and treatment of cancers |
US8795981B2 (en) | 2008-08-08 | 2014-08-05 | Molecular Devices, Llc | Cell detection |
MX2011001409A (en) | 2008-08-14 | 2011-03-29 | Cephalon Australia Pty Ltd | Anti-il-12/il-23 antibodies. |
WO2010027981A1 (en) | 2008-09-03 | 2010-03-11 | Genentech, Inc. | Multispecific antibodies |
US8163497B2 (en) | 2008-09-07 | 2012-04-24 | Glyconex Inc. | Anti-extended type I glycosphingolipid antibody, derivatives thereof and use |
CA2735716A1 (en) | 2008-09-12 | 2010-03-18 | Dako Denmark A/S | Prostate cancer biomarker |
US8417011B2 (en) | 2008-09-18 | 2013-04-09 | Molecular Devices (New Milton) Ltd. | Colony detection |
EP2352760A2 (en) * | 2008-09-22 | 2011-08-10 | Calmune Corporation | Methods and vectors for display of 2g12-derived domain exchanged antibodies |
US20100081575A1 (en) * | 2008-09-22 | 2010-04-01 | Robert Anthony Williamson | Methods for creating diversity in libraries and libraries, display vectors and methods, and displayed molecules |
WO2010034779A2 (en) | 2008-09-24 | 2010-04-01 | The Provost, Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth Near Dublin | Composition and method for treatment of preterm labor |
BRPI0918947A2 (en) | 2008-09-26 | 2015-12-01 | Ucb Pharma Sa | antibody fusion protein |
US9005963B2 (en) | 2008-10-14 | 2015-04-14 | Ablynx N.V. | Amino acid sequences directed against cellular receptors for viruses and bacteria |
MX345226B (en) | 2008-10-29 | 2017-01-20 | Ablynx Nv | Formulations of single domain antigen binding molecules. |
CA2739352C (en) | 2008-10-29 | 2021-07-13 | Wyeth Llc | Methods for purification of single domain antigen binding molecules |
WO2010054007A1 (en) | 2008-11-07 | 2010-05-14 | Fabrus Llc | Combinatorial antibody libraries and uses thereof |
CN102307902B (en) | 2008-12-10 | 2016-02-24 | 埃博灵克斯股份有限公司 | Be used for the treatment of and the aminoacid sequence for angiogenin/Tie system of relevant disease and illness occur to blood vessel and comprises its polypeptide |
US8775090B2 (en) | 2008-12-12 | 2014-07-08 | Medimmune, Llc | Crystals and structure of a human IgG Fc variant with enhanced FcRn binding |
ES2729320T3 (en) | 2008-12-19 | 2019-10-31 | Ablynx Nv | Genetic immunization to produce immunoglobulins against cell-associated antigens such as P2X7, CXCR7 or CXCR4 |
UY32341A (en) | 2008-12-19 | 2010-07-30 | Glaxo Group Ltd | NEW ANTIGEN UNION PROTEINS |
US20120003235A1 (en) | 2008-12-31 | 2012-01-05 | Biogen Idec Ma Inc. | Anti-lymphotoxin antibodies |
US20120058131A1 (en) | 2009-01-21 | 2012-03-08 | Oxford Biotherapeutics Ltd | Pta089 protein |
US20100260752A1 (en) | 2009-01-23 | 2010-10-14 | Biosynexus Incorporated | Opsonic and protective antibodies specific for lipoteichoic acid of gram positive bacteria |
JP2012516153A (en) | 2009-01-29 | 2012-07-19 | アボット・ラボラトリーズ | IL-1 binding protein |
EP2219029A1 (en) | 2009-01-30 | 2010-08-18 | Sanofi-Aventis | Test systems, methods and uses involving AS160 protein |
EP2398504B1 (en) | 2009-02-17 | 2018-11-28 | Cornell Research Foundation, Inc. | Methods and kits for diagnosis of cancer and prediction of therapeutic value |
JP5721639B2 (en) | 2009-02-19 | 2015-05-20 | ダコ・デンマーク・エー/エス | Methods and compounds for detection of molecular targets |
US8030026B2 (en) | 2009-02-24 | 2011-10-04 | Abbott Laboratories | Antibodies to troponin I and methods of use thereof |
US8420084B2 (en) | 2009-03-05 | 2013-04-16 | Medarex, Inc. | Fully human antibodies specific to CADM1 |
WO2010100437A2 (en) | 2009-03-05 | 2010-09-10 | University Of Sheffield | Production of protein |
US10005830B2 (en) | 2009-03-05 | 2018-06-26 | Ablynx N.V. | Antigen binding dimer-complexes, methods of making/avoiding and uses thereof |
RU2015132478A (en) | 2009-03-05 | 2015-12-10 | Эббви Инк. | BINDING IL-17 PROTEINS |
US8283162B2 (en) | 2009-03-10 | 2012-10-09 | Abbott Laboratories | Antibodies relating to PIVKAII and uses thereof |
SI3260136T1 (en) | 2009-03-17 | 2021-05-31 | Theraclone Sciences, Inc. | Human immunodeficiency virus (hiv) -neutralizing antibodies |
GB0905023D0 (en) | 2009-03-24 | 2009-05-06 | Univ Erasmus Medical Ct | Binding molecules |
PT2417156E (en) | 2009-04-07 | 2015-04-29 | Roche Glycart Ag | Trivalent, bispecific antibodies |
EP3461844A3 (en) | 2009-04-10 | 2019-05-15 | Ablynx N.V. | Improved amino acid sequences directed against il-6r and polypeptides comprising the same for the treatment of il-6r related diseases and disorders |
WO2010124163A2 (en) * | 2009-04-23 | 2010-10-28 | Theraclone Sciences, Inc. | Granulocyte-macrophage colony-stimulating factor (gm-csf) neutralizing antibodies |
DK2424889T3 (en) | 2009-04-30 | 2015-11-02 | Ablynx Nv | Process for the preparation of domain antibodies |
BRPI1011195B1 (en) * | 2009-05-20 | 2020-10-13 | Novimmune S.A | methods to produce a collection of nucleic acids |
JP5804521B2 (en) | 2009-05-29 | 2015-11-04 | モルフォシス・アー・ゲー | Collection and its usage |
EP2438087B1 (en) | 2009-06-05 | 2017-05-10 | Ablynx N.V. | Trivalent anti human respiratory syncytial virus (hrsv) nanobody constructs for the prevention and/or treatment of respiratory tract infections |
US9676845B2 (en) | 2009-06-16 | 2017-06-13 | Hoffmann-La Roche, Inc. | Bispecific antigen binding proteins |
EP2448972A4 (en) * | 2009-06-30 | 2012-11-28 | Res Dev Foundation | Immunoglobulin fc polypeptides |
EP2448966B1 (en) | 2009-07-03 | 2018-11-14 | Avipep Pty Ltd | Immuno-conjugates and methods for producing them |
IE20090514A1 (en) | 2009-07-06 | 2011-02-16 | Opsona Therapeutics Ltd | Humanised antibodies and uses therof |
ES2804450T3 (en) | 2009-07-10 | 2021-02-08 | Ablynx Nv | Method for producing variable domains |
WO2011014457A1 (en) | 2009-07-27 | 2011-02-03 | Genentech, Inc. | Combination treatments |
CN102741288B (en) | 2009-08-29 | 2015-08-19 | Abbvie公司 | DLL4 associated proteins is used in treatment |
WO2011026017A1 (en) | 2009-08-31 | 2011-03-03 | Abbott Laboratories | Biomarkers for prediction of major adverse cardiac events and uses thereof |
EP2293072A1 (en) | 2009-08-31 | 2011-03-09 | Sanofi-Aventis | Use of cathepsin H |
JP2013503607A (en) | 2009-09-01 | 2013-02-04 | アボット・ラボラトリーズ | Dual variable domain immunoglobulins and uses thereof |
EP2473522B1 (en) | 2009-09-02 | 2016-08-17 | Genentech, Inc. | Mutant smoothened and methods of using the same |
AU2010289677B2 (en) | 2009-09-03 | 2014-07-31 | Merck Sharp & Dohme Llc | Anti-GITR antibodies |
PL2473528T3 (en) | 2009-09-03 | 2015-05-29 | Ablynx Nv | Stable formulations of polypeptides and uses thereof |
WO2011032181A2 (en) * | 2009-09-14 | 2011-03-17 | Dyax Corp. | Libraries of genetic packages comprising novel hc cdr3 designs |
EP2477654A4 (en) | 2009-09-14 | 2013-01-23 | Abbott Lab | Methods for treating psoriasis |
US20120302737A1 (en) | 2009-09-16 | 2012-11-29 | Genentech, Inc. | Coiled coil and/or tether containing protein complexes and uses thereof |
US20110189183A1 (en) | 2009-09-18 | 2011-08-04 | Robert Anthony Williamson | Antibodies against candida, collections thereof and methods of use |
GB201005063D0 (en) | 2010-03-25 | 2010-05-12 | Ucb Pharma Sa | Biological products |
US8568726B2 (en) | 2009-10-06 | 2013-10-29 | Medimmune Limited | RSV specific binding molecule |
US8518405B2 (en) | 2009-10-08 | 2013-08-27 | The University Of North Carolina At Charlotte | Tumor specific antibodies and uses therefor |
TW201117824A (en) | 2009-10-12 | 2011-06-01 | Amgen Inc | Use of IL-17 receptor a antigen binding proteins |
US20120231004A1 (en) | 2009-10-13 | 2012-09-13 | Oxford Biotherapeutic Ltd. | Antibodies |
AR078651A1 (en) | 2009-10-15 | 2011-11-23 | Abbott Lab | IMMUNOGLOBULINS WITH DUAL VARIABLE DOMAIN AND USES OF THE SAME |
WO2011046457A1 (en) | 2009-10-16 | 2011-04-21 | Auckland Uniservices Limited | Anti-neoplastic uses of artemin antagonists |
CN104714007B (en) | 2009-10-20 | 2017-04-12 | 丹麦达科有限公司 | Immunochemical detection of single target entities |
UY32979A (en) | 2009-10-28 | 2011-02-28 | Abbott Lab | IMMUNOGLOBULINS WITH DUAL VARIABLE DOMAIN AND USES OF THE SAME |
JO3437B1 (en) | 2009-10-30 | 2019-10-20 | Esai R & D Man Co Ltd | Improved anti human Fraktalkine antibodies and uses thereof |
TW201121568A (en) | 2009-10-31 | 2011-07-01 | Abbott Lab | Antibodies to receptor for advanced glycation end products (RAGE) and uses thereof |
US20120282177A1 (en) | 2009-11-02 | 2012-11-08 | Christian Rohlff | ROR1 as Therapeutic and Diagnostic Target |
EP2496600A1 (en) | 2009-11-04 | 2012-09-12 | Fabrus LLC | Methods for affinity maturation-based antibody optimization |
PE20121646A1 (en) | 2009-11-04 | 2012-12-02 | Merck Sharp & Dohme | ANTI-TSLP ANTIBODY MODIFIED BY GENETIC ENGINEERING TECHNIQUES |
US20110165648A1 (en) | 2009-11-04 | 2011-07-07 | Menno Van Lookeren Campagne | Co-crystal structure of factor D and anti-factor D antibody |
CN102640001A (en) | 2009-11-05 | 2012-08-15 | 诺瓦提斯公司 | Biomarkers predictive of progression of fibrosis |
CA2780024A1 (en) | 2009-11-11 | 2011-05-19 | Gentian As | Immunoassay for assessing related analytes of different origin |
EP2507262A1 (en) | 2009-11-30 | 2012-10-10 | Ablynx N.V. | Improved amino acid sequences directed against human respiratory syncytial virus (hrsv) and polypeptides comprising the same for the prevention and/or treatment of respiratory tract infections |
CA3040276A1 (en) | 2009-12-02 | 2011-06-09 | Imaginab, Inc. | J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen (psma) and methods for their use |
EP2510001B1 (en) | 2009-12-08 | 2015-12-02 | AbbVie Deutschland GmbH & Co KG | Monoclonal antibodies against the rgm a protein for use in the treatment of retinal nerve fiber layer degeneration |
WO2011071577A1 (en) | 2009-12-11 | 2011-06-16 | Genentech, Inc. | Anti-vegf-c antibodies and methods using same |
EP3309176A1 (en) | 2009-12-14 | 2018-04-18 | Ablynx N.V. | Immunoglobulin single variable domain antibodies against ox40l, constructs and therapeutic use |
US20110226650A1 (en) | 2009-12-21 | 2011-09-22 | Genentech, Inc. | Antibody formulation |
CA2784385A1 (en) | 2009-12-23 | 2011-06-30 | Genentech, Inc. | Anti-bv8 antibodies and uses thereof |
CA2784610C (en) | 2009-12-23 | 2020-07-14 | Avipep Pty Ltd | Immuno-conjugates and methods for producing them |
WO2011083140A1 (en) | 2010-01-08 | 2011-07-14 | Ablynx Nv | Immunoglobulin single variable domain directed against human cxcr4 |
TWI513466B (en) | 2010-01-20 | 2015-12-21 | Boehringer Ingelheim Int | Anticoagulant antidotes |
US8940303B2 (en) | 2010-01-28 | 2015-01-27 | Glaxo Group Limited | CD127 binding proteins |
EP2354159A1 (en) | 2010-02-05 | 2011-08-10 | RWTH Aachen | CCL17 inhibitors for use in T helper cell-driven diseases |
US9120855B2 (en) | 2010-02-10 | 2015-09-01 | Novartis Ag | Biologic compounds directed against death receptor 5 |
MX2012009318A (en) | 2010-02-10 | 2012-09-07 | Novartis Ag | Methods and compounds for muscle growth. |
SI2533761T1 (en) | 2010-02-11 | 2019-08-30 | Ablynx N.V. | Methods and compositions for the preparation of aerosols |
MY160556A (en) | 2010-02-18 | 2017-03-15 | Genentech Inc | Neuregulin antagonists and use thereof in treating cancer |
RU2016146198A (en) | 2010-03-02 | 2018-12-19 | Эббви Инк. | THERAPEUTIC DLL4-BINDING PROTEINS |
UA108227C2 (en) | 2010-03-03 | 2015-04-10 | ANTIGENCY PROTEIN | |
GB201003701D0 (en) | 2010-03-05 | 2010-04-21 | Cilian Ag | System for the expression of a protein |
BR112012023895A2 (en) | 2010-03-17 | 2016-11-29 | Abbott Res Bv | anti nerve growth factor (ngf) compositions |
CA2791991A1 (en) | 2010-03-24 | 2011-09-29 | Genentech, Inc. | Anti-lrp6 antibodies |
US8937164B2 (en) | 2010-03-26 | 2015-01-20 | Ablynx N.V. | Biological materials related to CXCR7 |
TW201138821A (en) | 2010-03-26 | 2011-11-16 | Roche Glycart Ag | Bispecific antibodies |
CA2796339C (en) | 2010-04-15 | 2020-03-31 | Abbott Laboratories | Amyloid-beta binding proteins |
AU2011249783B9 (en) | 2010-05-06 | 2014-11-06 | Novartis Ag | Compositions and methods of use for therapeutic low density lipoprotein -related protein 6 (LRP6) antibodies |
US9290573B2 (en) | 2010-05-06 | 2016-03-22 | Novartis Ag | Therapeutic low density lipoprotein-related protein 6 (LRP6) multivalent antibodies |
ES2635594T3 (en) | 2010-05-14 | 2017-10-04 | Abbvie Inc. | IL-1 binding proteins |
JP6120767B2 (en) | 2010-05-20 | 2017-04-26 | アブリンクス ナームローゼ・フェノーツハップAblynx NV | Biological materials related to HER3 |
EP2577309B1 (en) | 2010-05-25 | 2016-11-23 | Carnegie Mellon University | Targeted probes of cellular physiology |
WO2011147834A1 (en) | 2010-05-26 | 2011-12-01 | Roche Glycart Ag | Antibodies against cd19 and uses thereof |
TW201210612A (en) | 2010-06-03 | 2012-03-16 | Glaxo Group Ltd | Humanised antigen binding proteins |
RU2613886C2 (en) | 2010-06-03 | 2017-03-21 | Дженентек, Инк. | Antibodies and immunoconjugates rendered by immuno-positron emission tomography, methods of application |
WO2011161545A2 (en) | 2010-06-04 | 2011-12-29 | The Netherlands Cancer Institute | Non-hydrolyzable protein conjugates, methods and compositions related thereto |
WO2011158019A1 (en) | 2010-06-16 | 2011-12-22 | Adjuvantix Limited | Polypeptide vaccine |
BR112012027995A2 (en) | 2010-06-18 | 2017-01-10 | Genentech Inc | antibody and isolated nucleic acid, host cell, method of producing an antibody, immunoconjugate, pharmaceutical formulation, use of the antibody, method of treating an individual with cancer, an individual having an immune disorder, inhibiting angiogenesis and inhibiting the constitutive activation of axl |
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 |
WO2011161263A1 (en) | 2010-06-25 | 2011-12-29 | Ablynx Nv | Pharmaceutical compositions for cutaneous administration |
WO2012006500A2 (en) | 2010-07-08 | 2012-01-12 | Abbott Laboratories | Monoclonal antibodies against hepatitis c virus core protein |
UY33492A (en) | 2010-07-09 | 2012-01-31 | Abbott Lab | IMMUNOGLOBULINS WITH DUAL VARIABLE DOMAIN AND USES OF THE SAME |
TW201217526A (en) | 2010-07-09 | 2012-05-01 | Biogen Idec Hemophilia Inc | Chimeric clotting factors |
KR20130120439A (en) | 2010-07-09 | 2013-11-04 | 제넨테크, 인크. | Anti-neuropilin antibodies and methods of use |
US20120100166A1 (en) | 2010-07-15 | 2012-04-26 | Zyngenia, Inc. | Ang-2 Binding Complexes and Uses Thereof |
JP6266343B2 (en) | 2010-07-16 | 2018-01-24 | アディマブ, エルエルシー | Antibody library |
CA2805653A1 (en) | 2010-07-20 | 2012-01-26 | Cephalon Australia Pty Ltd | Anti-il-23 heterodimer specific antibodies |
WO2012010582A1 (en) | 2010-07-21 | 2012-01-26 | Roche Glycart Ag | Anti-cxcr5 antibodies and methods of use |
US9120862B2 (en) | 2010-07-26 | 2015-09-01 | Abbott Laboratories | Antibodies relating to PIVKA-II and uses thereof |
WO2012018771A1 (en) | 2010-08-03 | 2012-02-09 | Genentech, Inc. | Chronic lymphocytic leukemia (cll) biomarkers |
NZ607480A (en) | 2010-08-03 | 2014-10-31 | Abbott Lab | Dual variable domain immunoglobulins and uses thereof |
MX2013001305A (en) | 2010-08-05 | 2013-03-20 | Hoffmann La Roche | Anti-mhc antibody anti-viral cytokine fusion protein. |
JP2013537416A (en) | 2010-08-13 | 2013-10-03 | メディミューン リミテッド | Monomer polypeptide containing mutant Fc region and method of use |
BR112013002444A2 (en) | 2010-08-13 | 2016-05-24 | Roche Glycart Ag | isolated antibody, polynucleotide and polypeptide, composition, vector, host cell, antibody conjugate, pharmaceutical formulation, use of the antibody, methods of producing an antibody, treating an individual, inducing cell lysis of a tumor cell and diagnosing a disease in an individual |
CN105884898B (en) | 2010-08-13 | 2022-10-11 | 罗切格利卡特公司 | Anti-fibroblast activation protein antibodies and methods of use |
JP6147665B2 (en) | 2010-08-14 | 2017-06-14 | アッヴィ・インコーポレイテッド | Amyloid beta-binding protein |
WO2012022734A2 (en) | 2010-08-16 | 2012-02-23 | Medimmune Limited | Anti-icam-1 antibodies and methods of use |
JP6121903B2 (en) | 2010-08-19 | 2017-04-26 | ゾエティス・ベルジャム・エス・アー | Anti-NGF antibodies and uses thereof |
DK2606070T3 (en) | 2010-08-20 | 2017-03-27 | Novartis Ag | Antibodies for the epidermal growth factor receptor 3 (HER3) |
BR112013001847A2 (en) | 2010-08-24 | 2016-05-31 | Hoffmann La Roche | bispecific antibody, method of preparation of bispecific antibody, trivalent bispecific antibody, methods and pharmaceutical composition |
CA2805054A1 (en) | 2010-08-25 | 2012-03-01 | F. Hoffmann-La Roche Ag | Antibodies against il-18r1 and uses thereof |
EP2608803A4 (en) | 2010-08-26 | 2014-01-15 | Abbvie Inc | Dual variable domain immunoglobulins and uses thereof |
EP3264089A1 (en) | 2010-08-31 | 2018-01-03 | Genentech, Inc. | Biomarkers and methods of treatment |
PT3556396T (en) | 2010-08-31 | 2022-07-04 | Scripps Research Inst | Human immunodeficiency virus (hiv)-neutralizing antibodies |
KR20130096731A (en) | 2010-09-08 | 2013-08-30 | 할로자임, 아이엔씨 | Methods for assessing and identifying or evolving conditionally active therapeutic proteins |
RU2551963C2 (en) | 2010-09-09 | 2015-06-10 | Пфайзер Инк. | Molecules binding to 4-1bb |
WO2012038744A2 (en) | 2010-09-22 | 2012-03-29 | Genome Research Limited | Detecting mutations |
GB201016494D0 (en) | 2010-09-30 | 2010-11-17 | Queen Mary Innovation Ltd | Polypeptide |
US8497138B2 (en) | 2010-09-30 | 2013-07-30 | Genetix Limited | Method for cell selection |
EP2625197B1 (en) | 2010-10-05 | 2016-06-29 | Genentech, Inc. | Mutant smoothened and methods of using the same |
PL2632946T3 (en) | 2010-10-29 | 2018-06-29 | Ablynx N.V. | Method for the production of immunoglobulin single variable domains |
DK2635607T3 (en) | 2010-11-05 | 2019-11-18 | Zymeworks Inc | STABLE HETERODIMED ANTIBODY DESIGN WITH MUTATIONS IN THE FC DOMAIN |
ES2723775T3 (en) | 2010-11-08 | 2019-09-02 | Ablynx Nv | Polypeptides that bind to CXCR2 |
US10180426B2 (en) | 2010-11-08 | 2019-01-15 | Dako Denmark A/S | Quantification of single target molecules in histological samples |
EP2638070B1 (en) | 2010-11-10 | 2016-10-19 | F.Hoffmann-La Roche Ag | Methods and compositions for neural disease immunotherapy |
MX353795B (en) | 2010-11-19 | 2018-01-30 | Morphosys Ag | A collection and methods for its use. |
CA2817161C (en) | 2010-12-06 | 2019-04-02 | Dako Denmark A/S | Combined histological stain |
SG10201401746TA (en) | 2010-12-16 | 2014-10-30 | Genentech Inc | Diagnosis And Treatments Relating To TH2 Inhibition |
TW201238974A (en) | 2010-12-17 | 2012-10-01 | Sanofi Sa | MiRNAs in joint disease |
TW201239097A (en) | 2010-12-17 | 2012-10-01 | Sanofi Sa | MiRNAs in joint disease |
TW201241179A (en) | 2010-12-17 | 2012-10-16 | Sanofi Sa | MiRNAs in joint disease |
UY33807A (en) | 2010-12-17 | 2012-07-31 | Sanofi Sa | miRNAs as indicators of tissue status or of diseases such as osteoarthritis |
TWI477513B (en) | 2010-12-20 | 2015-03-21 | 建南德克公司 | Anti-mesothelin antibodies and immunoconjugates |
WO2012121775A2 (en) | 2010-12-21 | 2012-09-13 | Abbott Laboratories | Dual variable domain immunoglobulins and uses thereof |
WO2012088222A2 (en) | 2010-12-21 | 2012-06-28 | The University Of North Carolina At Chapel Hill | Methods and compositions for producing active vitamin k-dependent proteins |
TW201307388A (en) | 2010-12-21 | 2013-02-16 | Abbott Lab | IL-1 binding proteins |
SG191219A1 (en) | 2010-12-22 | 2013-07-31 | Genentech Inc | Anti-pcsk9 antibodies and methods of use |
WO2012085064A1 (en) | 2010-12-23 | 2012-06-28 | Roche Diagnostics Gmbh | Detection of a posttranslationally modified polypeptide by a bi-valent binding agent |
WO2012085113A1 (en) | 2010-12-23 | 2012-06-28 | Roche Diagnostics Gmbh | Binding agent |
WO2012085069A2 (en) | 2010-12-23 | 2012-06-28 | Roche Diagnostics Gmbh | Detection of a polypeptide dimer by a bivalent binding agent |
SG191153A1 (en) | 2010-12-23 | 2013-07-31 | Hoffmann La Roche | Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery |
EP2471554A1 (en) | 2010-12-28 | 2012-07-04 | Hexal AG | Pharmaceutical formulation comprising a biopharmaceutical drug |
EP2658971A1 (en) | 2010-12-28 | 2013-11-06 | XOMA Technology Ltd. | Cell surface display using pdz domains |
US8952132B2 (en) | 2011-02-07 | 2015-02-10 | Research Development Foundation | Engineered immunoglobulin FC polypeptides |
CA2824252A1 (en) | 2011-02-10 | 2012-08-16 | Roche Glycart Ag | Improved immunotherapy |
WO2012109624A2 (en) | 2011-02-11 | 2012-08-16 | Zyngenia, Inc. | Monovalent and multivalent multispecific complexes and uses thereof |
WO2012116926A1 (en) | 2011-02-28 | 2012-09-07 | F. Hoffmann-La Roche Ag | Antigen binding proteins |
KR101572338B1 (en) | 2011-02-28 | 2015-11-26 | 에프. 호프만-라 로슈 아게 | Monovalent antigen binding proteins |
US9624294B2 (en) | 2011-03-14 | 2017-04-18 | Cellmid Limited | Antibody recognizing N-domain of midkine |
JP2014510730A (en) | 2011-03-16 | 2014-05-01 | サノフイ | Use of dual V region antibody-like proteins |
EP2691415B1 (en) | 2011-03-28 | 2018-07-11 | Ablynx N.V. | Method for producing solid formulations comprising immunoglobulin single variable domains |
US20150158948A9 (en) | 2011-03-28 | 2015-06-11 | Francis Descamps | Bispecific anti-cxcr7 immunoglobulin single variable domains |
AU2012234335B2 (en) | 2011-03-29 | 2016-09-01 | Roche Glycart Ag | Antibody Fc variants |
KR20140009437A (en) | 2011-03-30 | 2014-01-22 | 베링거 인겔하임 인터내셔날 게엠베하 | Anticoagulant antidotes |
CA2828890A1 (en) | 2011-04-07 | 2012-10-11 | Genentech, Inc. | Anti-fgfr4 antibodies and methods of use |
EP2511293A1 (en) | 2011-04-13 | 2012-10-17 | LEK Pharmaceuticals d.d. | A method for controlling the main complex N-glycan structures and the acidic variants and variability in bioprocesses producing recombinant proteins |
CA2832907C (en) | 2011-04-19 | 2020-07-14 | Dako Denmark A/S | New method for enzyme-mediated signal amplification |
RU2013150331A (en) | 2011-04-20 | 2015-05-27 | Рош Гликарт Аг | METHOD AND DEVICES FOR A pH-DEPENDENT PASSAGE OF A HEMATOENCEPHALIC BARRIER |
EP2699597B1 (en) | 2011-04-21 | 2016-06-01 | Garvan Institute of Medical Research | Modified variable domain molecules and methods for producing and using them b |
EP2518157A1 (en) | 2011-04-26 | 2012-10-31 | Sanofi | Test Systems and methods for identifying a compound altering cellular DDR activity |
US9062106B2 (en) | 2011-04-27 | 2015-06-23 | 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 |
UA117218C2 (en) | 2011-05-05 | 2018-07-10 | Мерк Патент Гмбх | Amino acid sequences directed against il-17a, il-17f and/or il17-a/f and polypeptides comprising the same |
DK3498732T3 (en) | 2011-05-06 | 2022-01-17 | Zoetis Services Llc | ANTIBODIES TO ANTI-NERVE GROWTH FACTOR AND METHODS OF MANUFACTURE AND USE |
GB201114858D0 (en) | 2011-08-29 | 2011-10-12 | Nvip Pty Ltd | Anti-nerve growth factor antibodies and methods of using the same |
KR101833465B1 (en) | 2011-05-06 | 2018-02-28 | 넥스베트 오스트레일리아 피티와이 리미티드 | Anti-nerve growth factor antibodies and methods of preparing and using the same |
CA2835094C (en) | 2011-05-06 | 2020-12-22 | David Gearing | Anti-nerve growth factor antibodies and methods of preparing and using the same |
SG10201500953YA (en) | 2011-05-06 | 2015-04-29 | Nvip Pty Ltd | Anti-nerve growth factor antibodies and methods of preparing and using the same |
WO2012152823A1 (en) | 2011-05-09 | 2012-11-15 | Ablynx Nv | Method for the production of immunoglobulin single variable domains |
MX2013013054A (en) | 2011-05-12 | 2014-02-20 | Genentech Inc | Multiple reaction monitoring lc-ms/ms method to detect therapeutic antibodies in animal samples using framework signature peptides. |
CA2834879C (en) | 2011-05-16 | 2019-10-22 | Genentech, Inc. | Fgfr1 agonists and methods of use |
IL295205A (en) | 2011-05-17 | 2022-10-01 | Univ Rockefeller | Human immunodeficiency virus neutralizing antibodies and methods of use thereof |
CN103857699B (en) | 2011-05-24 | 2016-08-31 | 泽恩格尼亚股份有限公司 | Multivalence and unit price polyspecific complex and application thereof |
CA2835340A1 (en) | 2011-05-27 | 2012-12-06 | Ablynx Nv | Inhibition of bone resorption with rankl binding peptides |
US9580480B2 (en) | 2011-05-31 | 2017-02-28 | Massachusetts Institute Of Technology | Cell-directed synthesis of multifunctional nanopatterns and nanomaterials |
WO2012170807A2 (en) | 2011-06-10 | 2012-12-13 | Medimmune, Llc | Anti-pseudomonas psl binding molecules and uses thereof |
KR101822702B1 (en) | 2011-06-13 | 2018-01-26 | 씨에스엘 리미티드 | Antibodies against g-csfr and uses thereof |
JP5984919B2 (en) | 2011-06-15 | 2016-09-06 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Anti-human EPO receptor antibody and method of use |
EP2537532A1 (en) | 2011-06-22 | 2012-12-26 | J. Stefan Institute | Cathepsin-binding compounds bound to a nanodevice and their diagnostic and therapeutic use |
EP4350345A2 (en) | 2011-06-23 | 2024-04-10 | Ablynx N.V. | Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobin single variable domains |
CN104271598A (en) | 2011-06-23 | 2015-01-07 | 埃博灵克斯股份有限公司 | Immunoglobulin single variable domains directed against IgE |
CA3142288A1 (en) | 2011-06-23 | 2012-12-27 | Ablynx Nv | Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains |
SI2723769T2 (en) | 2011-06-23 | 2022-09-30 | Ablynx Nv | Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains |
WO2013001369A2 (en) | 2011-06-28 | 2013-01-03 | Oxford Biotherapeutics Ltd. | Therapeutic and diagnostic target |
WO2013003625A2 (en) | 2011-06-28 | 2013-01-03 | Oxford Biotherapeutics Ltd. | Antibodies |
TW201306866A (en) | 2011-06-30 | 2013-02-16 | Genentech Inc | Anti-c-met antibody formulations |
AU2012283039A1 (en) | 2011-07-13 | 2014-01-30 | Abbvie Inc. | Methods and compositions for treating asthma using anti-IL-13 antibodies |
WO2013012733A1 (en) | 2011-07-15 | 2013-01-24 | Biogen Idec Ma Inc. | Heterodimeric fc regions, binding molecules comprising same, and methods relating thereto |
WO2013016220A1 (en) | 2011-07-22 | 2013-01-31 | Amgen Inc. | Il-17 receptor a is required for il-17c biology |
BR112014003431A2 (en) | 2011-08-17 | 2017-06-13 | Genentech Inc | antibody, nucleic acid, host cell, method of producing an antibody, immunoconjugate, pharmaceutical formulation, pharmaceutical agent, use of the antibody, method of treating an individual who has cancer, and time-lapse method for tumor recurrence |
MX2014002053A (en) | 2011-08-23 | 2014-04-25 | Roche Glycart Ag | Anti-mcsp antibodies. |
EP3321286B1 (en) | 2011-08-23 | 2021-01-06 | Roche Glycart AG | Bispecific t cell activating antigen binding molecules |
BR112014004168A2 (en) | 2011-08-23 | 2017-12-12 | Roche Glycart Ag | bispecific antibody, pharmaceutical composition, use of bispecific antibody, prokaryotic or eukaryotic host cell, antibody production method and invention |
SG2014008577A (en) | 2011-08-23 | 2014-04-28 | Roche Glycart Ag | Bispecific antigen binding molecules |
WO2013026837A1 (en) | 2011-08-23 | 2013-02-28 | Roche Glycart Ag | Bispecific t cell activating antigen binding molecules |
SG11201400222RA (en) | 2011-08-30 | 2014-03-28 | Nvip Pty Ltd | Caninised tumour necrosis factor antibodies and methods of using the same |
US8822651B2 (en) | 2011-08-30 | 2014-09-02 | Theraclone Sciences, Inc. | Human rhinovirus (HRV) antibodies |
CA2848074A1 (en) | 2011-09-09 | 2013-03-14 | Medimmune Limited | Anti-siglec-15 antibodies and uses thereof |
JP2014527180A (en) | 2011-09-14 | 2014-10-09 | アベテルノ リミテッド | Intracellular sorting |
KR20140068062A (en) | 2011-09-15 | 2014-06-05 | 제넨테크, 인크. | Methods of promoting differentiation |
RU2014114617A (en) | 2011-09-19 | 2015-10-27 | Дженентек, Инк. | COMBINED TREATMENTS CONTAINING C-MET ANTAGONISTS AND B-RAF ANTAGONISTS |
GB201116116D0 (en) | 2011-09-19 | 2011-11-02 | Univ York | Cell differentiation |
SG10201703771WA (en) | 2011-09-19 | 2017-06-29 | Axon Neuroscience Se | Protein-based therapy and diagnosis of tau-mediated pathology in alzheimer's disease |
JO3625B1 (en) | 2011-09-22 | 2020-08-27 | Amgen Inc | CD27L Antigen Binding Proteins |
EP2747782B1 (en) | 2011-09-23 | 2018-01-17 | Ablynx NV | Prolonged inhibition of interleukin-6 mediated signaling |
GB201116702D0 (en) | 2011-09-28 | 2011-11-09 | Procure Therapeutics Ltd | Cell surface markers |
AU2012315474B2 (en) | 2011-09-30 | 2017-10-26 | Teva Pharmaceuticals Australia Pty Ltd. | Antibodies against TL1a and uses thereof |
SG11201401287SA (en) | 2011-10-05 | 2014-05-29 | Genentech Inc | Methods of treating liver conditions using notch2 antagonists |
US9575073B2 (en) | 2011-10-10 | 2017-02-21 | Rutgers, The State University Of New Jersey | Detection of high-risk intraductal papillary mucinous neoplasm and pancreatic adenocarcinoma |
KR20140082796A (en) | 2011-10-14 | 2014-07-02 | 제넨테크, 인크. | ANTI-HtrA1 ANTIBODIES AND METHODS OF USE |
EP2766000A2 (en) | 2011-10-15 | 2014-08-20 | F.Hoffmann-La Roche Ag | Scd1 antagonists for treating cancer |
WO2013059531A1 (en) | 2011-10-20 | 2013-04-25 | Genentech, Inc. | Anti-gcgr antibodies and uses thereof |
WO2013063114A1 (en) | 2011-10-24 | 2013-05-02 | Abbvie Inc. | Immunobinders directed against tnf |
EP2771361A1 (en) | 2011-10-24 | 2014-09-03 | AbbVie Inc. | Bispecific immunobinders directed against tnf and il-17 |
TW201323440A (en) | 2011-10-24 | 2013-06-16 | Abbvie Inc | Immunobinders directed against sclerostin |
GB201118359D0 (en) | 2011-10-25 | 2011-12-07 | Univ Sheffield | Pulmonary hypertension |
AU2012328980A1 (en) | 2011-10-28 | 2014-04-24 | Genentech, Inc. | Therapeutic combinations and methods of treating melanoma |
EA201700111A1 (en) | 2011-10-28 | 2018-02-28 | Тева Фармасьютикал Австралия Пти Лтд | POLYPEPTIDE STRUCTURES AND THEIR APPLICATION |
CA2852709A1 (en) | 2011-10-28 | 2013-05-02 | Patrys Limited | Pat-lm1 epitopes and methods for using same |
GB201118840D0 (en) | 2011-11-01 | 2011-12-14 | Univ Sheffield | Pulmonary hypertension II |
ES2899956T3 (en) | 2011-11-04 | 2022-03-15 | Zymeworks Inc | Stable heterodimeric antibody design with mutations in the Fc domain |
JP2015502741A (en) | 2011-11-04 | 2015-01-29 | ノバルティス アーゲー | Low density lipoprotein related protein 6 (LRP6)-half-life extended construct |
EP2776466B1 (en) | 2011-11-11 | 2017-08-23 | UCB Biopharma SPRL | Albumin binding antibodies and binding fragments thereof |
RU2014124842A (en) | 2011-11-21 | 2015-12-27 | Дженентек, Инк. | CLEANING ANTI-C-MET ANTIBODIES |
BR112014012882A2 (en) | 2011-11-29 | 2017-06-13 | Genentech Inc | method, antibody, polynucleotide, host cell, hybridoma cell line, antibody use and kit |
AU2012346861A1 (en) | 2011-11-30 | 2014-06-19 | AbbVie Deutschland GmbH & Co. KG | Methods and compositions for determining responsiveness to treatment with a tnf-alpha inhibitor |
EA201491107A1 (en) | 2011-12-05 | 2014-11-28 | Новартис Аг | ANTIBODIES TO THE RECEPTOR EPIDERMAL GROWTH FACTOR 3 (HER3), DIRECTED TO DOMAIN II HER3 |
CN108341873B (en) | 2011-12-05 | 2022-03-25 | 诺华股份有限公司 | Antibodies to epidermal growth factor receptor 3(HER3) |
US20140335084A1 (en) | 2011-12-06 | 2014-11-13 | Hoffmann-La Roche Inc. | Antibody formulation |
EP2794878B1 (en) | 2011-12-22 | 2020-03-18 | F.Hoffmann-La Roche Ag | Expression vector organization, novel production cell generation methods and their use for the recombinant production of polypeptides |
SG11201403443WA (en) | 2011-12-22 | 2014-07-30 | Hoffmann La Roche | Expression vector element combinations, 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 |
EP2797955A2 (en) | 2011-12-30 | 2014-11-05 | AbbVie Inc. | Dual variable domain immunoglobulins against il-13 and/or il-17 |
US20150011431A1 (en) | 2012-01-09 | 2015-01-08 | The Scripps Research Institute | Humanized antibodies |
US20140050720A1 (en) | 2012-01-09 | 2014-02-20 | The Scripps Research Institute | Ultralong complementarity determining regions and uses thereof |
JP6247226B2 (en) | 2012-01-10 | 2017-12-13 | バイオジェン・エムエイ・インコーポレイテッドBiogen MA Inc. | Improved transport of therapeutic molecules across the blood brain barrier |
WO2013109856A2 (en) | 2012-01-18 | 2013-07-25 | Genentech, Inc. | Methods of using fgf19 modulators |
IN2014DN05885A (en) | 2012-01-18 | 2015-06-05 | Hoffmann La Roche | |
PL2807192T3 (en) | 2012-01-27 | 2019-02-28 | Abbvie Deutschland | Composition and method for diagnosis and treatment of diseases associated with neurite degeneration |
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 |
JP6486686B2 (en) | 2012-02-10 | 2019-03-20 | ジェネンテック, インコーポレイテッド | Single chain antibodies and other heteromultimers |
CN113398268A (en) | 2012-02-11 | 2021-09-17 | 霍夫曼-拉罗奇有限公司 | R-spondin translocations and methods of use thereof |
US20150018241A1 (en) | 2012-02-15 | 2015-01-15 | Hoffmann-La Roche Inc. | Fc-receptor based affinity chromatography |
GB201203071D0 (en) | 2012-02-22 | 2012-04-04 | Ucb Pharma Sa | Biological products |
GB201203587D0 (en) | 2012-03-01 | 2012-04-11 | Univ Warwick | Modified bacterial cell |
CA2860369A1 (en) | 2012-03-02 | 2013-09-06 | Roche Glycart Ag | Predicitive biomarker for cancer treatment with adcc enhanced antibodies |
US9592289B2 (en) | 2012-03-26 | 2017-03-14 | Sanofi | Stable IgG4 based binding agent formulations |
US10041950B2 (en) | 2012-03-27 | 2018-08-07 | Ventana Medical Systems, Inc. | Signaling conjugates and methods of use |
JP2015514710A (en) | 2012-03-27 | 2015-05-21 | ジェネンテック, インコーポレイテッド | Diagnosis and treatment of HER3 inhibitors |
AR090549A1 (en) | 2012-03-30 | 2014-11-19 | Genentech Inc | ANTI-LGR5 AND IMMUNOCATE PLAYERS |
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 |
AR090903A1 (en) | 2012-05-01 | 2014-12-17 | Genentech Inc | ANTI-PMEL ANTIBODIES AND IMMUNOCADES17 |
US9328174B2 (en) | 2012-05-09 | 2016-05-03 | Novartis Ag | Chemokine receptor binding polypeptides |
AU2013258834B2 (en) | 2012-05-10 | 2017-09-07 | Zymeworks Bc Inc. | Heteromultimer constructs of immunoglobulin heavy chains with mutations in the Fc domain |
WO2013170191A1 (en) | 2012-05-11 | 2013-11-14 | Genentech, Inc. | Methods of using antagonists of nad biosynthesis from nicotinamide |
WO2013173364A2 (en) | 2012-05-14 | 2013-11-21 | Biogen Idec Ma Inc. | Lingo-2 antagonists for treatment of conditions involving motor neurons |
UY34813A (en) | 2012-05-18 | 2013-11-29 | Amgen Inc | ANTIGEN UNION PROTEINS DIRECTED AGAINST ST2 RECEIVER |
EP2666786A1 (en) | 2012-05-21 | 2013-11-27 | PAION Deutschland GmbH | Immunotherapy for intracranial hemorrhage |
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 |
EP2855521A4 (en) | 2012-05-24 | 2016-03-02 | Mountgate Group Ltd | Compositions and methods related to prevention and treatment of rabies infection |
AR091462A1 (en) | 2012-06-15 | 2015-02-04 | Genentech Inc | ANTI-PCSK9 ANTIBODIES, FORMULATIONS, DOSAGE AND METHODS OF USE |
US9499634B2 (en) | 2012-06-25 | 2016-11-22 | Zymeworks Inc. | Process and methods for efficient manufacturing of highly pure asymmetric antibodies in mammalian cells |
RU2639287C2 (en) | 2012-06-27 | 2017-12-20 | Ф. Хоффманн-Ля Рош Аг | Method for selection and obtaining of highly selective and multispecific targeting groups with specified properties, including at least two different binding groups, and their applications |
WO2014001325A1 (en) | 2012-06-27 | 2014-01-03 | F. Hoffmann-La Roche Ag | Method for making antibody fc-region conjugates comprising at least one binding entity that specifically binds to a target and uses thereof |
CA2871386A1 (en) | 2012-06-27 | 2014-01-03 | F. Hoffmann-La Roche Ag | Method for the selection and production of tailor-made, selective and multi-specific therapeutic molecules comprising at least two different targeting entities and uses thereof |
PL2869837T3 (en) | 2012-07-04 | 2017-03-31 | F.Hoffmann-La Roche Ag | Anti-theophylline antibodies and methods of use |
JP6247287B2 (en) | 2012-07-04 | 2017-12-13 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Anti-biotin antibodies and methods of use |
AU2013285422B2 (en) | 2012-07-04 | 2017-04-27 | F. Hoffmann-La Roche Ag | Covalently linked antigen-antibody conjugates |
KR20150030693A (en) | 2012-07-05 | 2015-03-20 | 제넨테크, 인크. | Expression and secretion system |
IN2014DN10652A (en) | 2012-07-09 | 2015-09-11 | Genentech Inc | |
EA201590174A1 (en) | 2012-07-09 | 2015-09-30 | Дженентек, Инк. | IMMUNOCONGATES CONTAINING ANTI-CD22 ANTIBODIES |
KR20150030698A (en) | 2012-07-09 | 2015-03-20 | 제넨테크, 인크. | Immunoconjugates comprising anti-cd79b antibodies |
TW201408696A (en) | 2012-07-09 | 2014-03-01 | Genentech Inc | Anti-CD22 antibodies and immunoconjugates |
UY34905A (en) | 2012-07-12 | 2014-01-31 | Abbvie Inc | IL-1 UNION PROTEINS |
EP3495387B1 (en) | 2012-07-13 | 2021-09-01 | Roche Glycart AG | Bispecific anti-vegf/anti-ang-2 antibodies and their use in the treatment of ocular vascular diseases |
CN112587658A (en) | 2012-07-18 | 2021-04-02 | 博笛生物科技有限公司 | Targeted immunotherapy for cancer |
GB201213652D0 (en) | 2012-08-01 | 2012-09-12 | Oxford Biotherapeutics Ltd | Therapeutic and diagnostic target |
JP6290209B2 (en) | 2012-08-07 | 2018-03-07 | ロシュ グリクアート アーゲー | A composition comprising two antibodies engineered to have reduced and increased effector function. |
BR112015002085A2 (en) | 2012-08-08 | 2017-12-19 | Roche Glycart Ag | protein, polynucleotide, vector, host cell, method for producing protein, pharmaceutical composition, protein use, method of treatment and invention |
CN109705218B (en) | 2012-08-09 | 2022-07-19 | 罗切格利卡特公司 | ASGPR antibodies and uses thereof |
WO2014035475A1 (en) | 2012-09-02 | 2014-03-06 | Abbvie Inc. | Methods to control protein heterogeneity |
US9512214B2 (en) | 2012-09-02 | 2016-12-06 | Abbvie, Inc. | Methods to control protein heterogeneity |
KR20150064068A (en) | 2012-10-08 | 2015-06-10 | 로슈 글리카트 아게 | FC-FREE ANTIBODIES COMPRISING TWO Fab-FRAGMENTS AND METHODS OF USE |
DK2908912T3 (en) | 2012-10-18 | 2020-10-26 | Univ Rockefeller | WIDE NEUTRALIZING ANTI-HIV ANTIBODIES |
US9688741B2 (en) | 2012-10-23 | 2017-06-27 | Elastagen Pty Ltd | Elastic hydrogel |
SG11201503412RA (en) | 2012-11-01 | 2015-05-28 | Abbvie Inc | Anti-vegf/dll4 dual variable domain immunoglobulins and uses thereof |
EP2914621B1 (en) | 2012-11-05 | 2023-06-07 | Foundation Medicine, Inc. | Novel ntrk1 fusion molecules and uses thereof |
EP2917243B1 (en) | 2012-11-08 | 2018-03-14 | F.Hoffmann-La Roche Ag | Her3 antigen binding proteins binding to the beta-hairpin of her3 |
MA38176A1 (en) | 2012-11-13 | 2017-06-30 | Genentech Inc | Novel anti-haemagglutinin antibody, useful for the treatment, inhibition or prevention of viral influenza infection |
US9914785B2 (en) | 2012-11-28 | 2018-03-13 | Zymeworks Inc. | Engineered immunoglobulin heavy chain-light chain pairs and uses thereof |
TW201425336A (en) | 2012-12-07 | 2014-07-01 | Amgen Inc | BCMA antigen binding proteins |
KR102239867B1 (en) | 2012-12-10 | 2021-04-13 | 앨러간 파마슈티컬스 인터내셔널 리미티드 | Scalable three-dimensional elastic construct manufacturing |
WO2014100542A1 (en) | 2012-12-21 | 2014-06-26 | Abbvie, Inc. | High-throughput antibody humanization |
JP2016510319A (en) | 2012-12-28 | 2016-04-07 | アッヴィ・インコーポレイテッド | Multivalent binding protein composition |
EP3939614A1 (en) | 2013-01-18 | 2022-01-19 | Foundation Medicine, Inc. | Methods of treating cholangiocarcinoma |
EP2948177A1 (en) | 2013-01-22 | 2015-12-02 | AbbVie Inc. | Methods for optimizing domain stability of binding proteins |
WO2014114595A1 (en) | 2013-01-23 | 2014-07-31 | Roche Glycart Ag | Predictive biomarker for cancer treatment with adcc-enhanced antibodies |
WO2014116846A2 (en) | 2013-01-23 | 2014-07-31 | Abbvie, Inc. | Methods and compositions for modulating an immune response |
WO2014116749A1 (en) | 2013-01-23 | 2014-07-31 | Genentech, Inc. | Anti-hcv antibodies and methods of using thereof |
ES2728936T3 (en) | 2013-01-25 | 2019-10-29 | Amgen Inc | Antibodies directed against CDH19 for melanoma |
EP2951208B1 (en) | 2013-02-01 | 2019-11-13 | Kira Biotech Pty Limited | Anti-cd83 antibodies and use thereof |
SG11201505762XA (en) | 2013-02-07 | 2015-08-28 | Csl Ltd | Il-11r binding proteins and uses thereof |
US20140228875A1 (en) | 2013-02-08 | 2014-08-14 | Nidus Medical, Llc | Surgical device with integrated visualization and cauterization |
GB201302447D0 (en) | 2013-02-12 | 2013-03-27 | Oxford Biotherapeutics Ltd | Therapeutic and diagnostic target |
JP2016509045A (en) | 2013-02-22 | 2016-03-24 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | How to treat cancer and prevent drug resistance |
WO2014131711A1 (en) | 2013-02-26 | 2014-09-04 | Roche Glycart Ag | Bispecific t cell activating antigen binding molecules |
JP6499087B2 (en) | 2013-02-26 | 2019-04-10 | ロシュ グリクアート アーゲー | Bispecific T cell activation antigen binding molecule |
EP2961771B1 (en) | 2013-02-26 | 2020-01-01 | Roche Glycart AG | Bispecific t cell activating antigen binding molecules specific to cd3 and cea |
MX2015010789A (en) | 2013-02-26 | 2015-11-26 | Roche Glycart Ag | Anti-mcsp antibodies. |
CA2902068C (en) | 2013-02-28 | 2023-10-03 | Caprion Proteomics Inc. | Tuberculosis biomarkers and uses thereof |
JP2016510751A (en) | 2013-03-06 | 2016-04-11 | ジェネンテック, インコーポレイテッド | Methods of treating and preventing anticancer drug resistance |
EP2830651A4 (en) | 2013-03-12 | 2015-09-02 | Abbvie Inc | Human antibodies that bind human tnf-alpha and methods of preparing the same |
SG10201913932VA (en) | 2013-03-13 | 2020-03-30 | Genentech Inc | Antibody formulations |
SI2968467T1 (en) | 2013-03-13 | 2020-11-30 | F. Hoffmann-La Roche Ag | Formulations with reduced oxidation |
AR095398A1 (en) | 2013-03-13 | 2015-10-14 | Genentech Inc | FORMULATIONS WITH REDUCED OXIDATION |
CN104968362B (en) | 2013-03-13 | 2018-12-14 | 霍夫曼-拉罗奇有限公司 | Aoxidize reduced preparaton |
WO2014143622A1 (en) | 2013-03-13 | 2014-09-18 | Seattle Genetics, Inc. | ACTIVATED CARBON FILTRATION FOR PURIFICATION OF BENZODIAZEPINE ADCs |
AR095399A1 (en) | 2013-03-13 | 2015-10-14 | Genentech Inc | FORMULATIONS WITH REDUCED OXIDATION, METHOD |
WO2014159239A2 (en) | 2013-03-14 | 2014-10-02 | Novartis Ag | Antibodies against notch 3 |
CA2906421C (en) | 2013-03-14 | 2022-08-16 | George J. Dawson | Hcv antigen-antibody combination assay and methods and compositions for use therein |
US8921526B2 (en) | 2013-03-14 | 2014-12-30 | Abbvie, Inc. | Mutated anti-TNFα antibodies and methods of their use |
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 |
WO2014152358A2 (en) | 2013-03-14 | 2014-09-25 | Genentech, Inc. | Combinations of a mek inhibitor compound with an her3/egfr inhibitor compound and methods of use |
US9562099B2 (en) | 2013-03-14 | 2017-02-07 | Genentech, Inc. | Anti-B7-H4 antibodies and immunoconjugates |
WO2014151878A2 (en) | 2013-03-14 | 2014-09-25 | Abbvie Inc. | Methods for modulating protein glycosylation profiles of recombinant protein therapeutics using monosaccharides and oligosacharides |
CN105307683A (en) | 2013-03-14 | 2016-02-03 | 基因泰克公司 | Methods of treating cancer and preventing cancer drug resistance |
CN105378099B (en) | 2013-03-14 | 2021-05-11 | 雅培制药有限公司 | HCV core lipid binding domain monoclonal antibodies |
KR20150127199A (en) | 2013-03-14 | 2015-11-16 | 제넨테크, 인크. | Anti-b7-h4 antibodies and immunoconjugates |
US9790478B2 (en) | 2013-03-14 | 2017-10-17 | Abbott Laboratories | HCV NS3 recombinant antigens and mutants thereof for improved antibody detection |
US20140286969A1 (en) | 2013-03-15 | 2014-09-25 | Abbvie Inc. | Anti-egfr antibody drug conjugate formulations |
CN105143264A (en) | 2013-03-15 | 2015-12-09 | 豪夫迈·罗氏有限公司 | Compositions and methods for diagnosis and treatment of hepatic cancers |
EP2970484B2 (en) | 2013-03-15 | 2022-09-21 | Amgen Inc. | Heterodimeric bispecific antibodies |
JP2016520528A (en) | 2013-03-15 | 2016-07-14 | ジェネンテック, インコーポレイテッド | Cancer treatment and anticancer drug resistance prevention method |
EP2968589A1 (en) | 2013-03-15 | 2016-01-20 | AbbVie Inc. | Antibody drug conjugate (adc) purification |
KR102202476B1 (en) | 2013-03-15 | 2021-01-12 | 제넨테크, 인크. | Cell culture media and methods of antibody production |
WO2014141192A1 (en) | 2013-03-15 | 2014-09-18 | Erasmus University Medical Center | Generation of heavy chain-only antibodies |
EP3424530A1 (en) | 2013-03-15 | 2019-01-09 | Zyngenia, Inc. | Multivalent and monovalent multispecific complexes and their uses |
WO2014151006A2 (en) | 2013-03-15 | 2014-09-25 | Genentech, Inc. | Biomarkers and methods of treating pd-1 and pd-l1 related conditions |
MX2015013166A (en) | 2013-03-15 | 2015-12-11 | Abbvie Inc | Dual specific binding proteins directed against il-1 beta and il-17. |
BR112015021993A8 (en) | 2013-03-15 | 2019-12-03 | Genentech Inc | polypeptide, methods for producing it, methods for culturing a cell, pharmaceutical composition, kit, and cell culture medium |
AR095517A1 (en) | 2013-03-15 | 2015-10-21 | Genentech Inc | ANTIBODIES AGAINST THE CHEMIOATRAYENT RECEIVER EXPRESSED IN T HELPER 2 CELLS (ANTI-CRTh2) AND METHODS OF USE |
US20140302037A1 (en) | 2013-03-15 | 2014-10-09 | Amgen Inc. | BISPECIFIC-Fc MOLECULES |
EP2970452A2 (en) | 2013-03-15 | 2016-01-20 | AC Immune S.A. | Anti-tau antibodies and methods of use |
GB201306589D0 (en) | 2013-04-11 | 2013-05-29 | Abeterno Ltd | Live cell imaging |
US11117975B2 (en) | 2013-04-29 | 2021-09-14 | Teva Pharmaceuticals Australia Pty Ltd | Anti-CD38 antibodies and fusions to attenuated interferon alpha-2B |
CA2909952C (en) | 2013-04-29 | 2021-10-12 | Teva Pharmaceuticals Australia Pty Ltd. | Anti-cd38 antibodies and fusions to attenuated interferon alpha-2b |
HUE047580T2 (en) | 2013-04-29 | 2020-05-28 | Agrosavfe Nv | Agrochemical compositions comprising antibodies binding to sphingolipids |
PL3594240T3 (en) | 2013-05-20 | 2024-04-02 | F. Hoffmann-La Roche Ag | Anti-transferrin receptor antibodies and methods of use |
NZ753995A (en) | 2013-05-30 | 2022-07-01 | Kiniksa Pharmaceuticals Ltd | Oncostatin m receptor antigen binding proteins |
US20160168231A1 (en) | 2013-07-18 | 2016-06-16 | Fabrus, Inc. | Antibodies with ultralong complementarity determining regions |
CA2918370A1 (en) | 2013-07-18 | 2015-01-22 | Fabrus, Inc. | Humanized antibodies with ultralong complementarity determining regions |
WO2015011660A1 (en) | 2013-07-23 | 2015-01-29 | Biocon Limited | Methods for controlling fucosylation levels in proteins |
CN113683695A (en) | 2013-08-02 | 2021-11-23 | 辉瑞公司 | anti-CXCR 4 antibodies and antibody-drug conjugates |
CA2920312C (en) | 2013-08-13 | 2022-07-19 | Elastagen Pty Ltd | Regeneration of damaged tissue |
RU2016109247A (en) | 2013-09-17 | 2017-10-19 | Дженентек, Инк. | WAYS OF APPLICATION OF ANTIBODIES TO LGR5 |
HUE047194T2 (en) | 2013-09-27 | 2020-04-28 | Hoffmann La Roche | Anti-pdl1 antibody formulations |
WO2015050959A1 (en) | 2013-10-01 | 2015-04-09 | Yale University | Anti-kit antibodies and methods of use thereof |
SG11201602522VA (en) | 2013-10-02 | 2016-04-28 | Medimmune Llc | Neutralizing anti-influenza a antibodies and uses thereof |
WO2015051293A2 (en) | 2013-10-04 | 2015-04-09 | Abbvie, Inc. | Use of metal ions for modulation of protein glycosylation profiles of recombinant proteins |
JP2016537965A (en) | 2013-10-11 | 2016-12-08 | ジェネンテック, インコーポレイテッド | NSP4 inhibitors and methods of use |
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 |
US9085618B2 (en) | 2013-10-18 | 2015-07-21 | Abbvie, Inc. | Low acidic species compositions and methods for producing and using the same |
US9181337B2 (en) | 2013-10-18 | 2015-11-10 | Abbvie, Inc. | Modulated lysine variant species compositions and methods for producing and using the same |
US8946395B1 (en) | 2013-10-18 | 2015-02-03 | Abbvie Inc. | Purification of proteins using hydrophobic interaction chromatography |
KR20160068802A (en) | 2013-10-23 | 2016-06-15 | 제넨테크, 인크. | Methods of diagnosing and treating eosinophilic disorders |
WO2015069459A1 (en) | 2013-11-05 | 2015-05-14 | Novartis Ag | Organic compounds |
WO2015073884A2 (en) | 2013-11-15 | 2015-05-21 | Abbvie, Inc. | Glycoengineered binding protein compositions |
LT3071597T (en) | 2013-11-21 | 2020-10-12 | F. Hoffmann-La Roche Ag | Anti-alpha-synuclein antibodies and methods of use |
EP3074424A4 (en) | 2013-11-27 | 2017-06-14 | Zymeworks Inc. | Bispecific antigen-binding constructs targeting her2 |
US9803008B2 (en) | 2013-11-28 | 2017-10-31 | Csl Limited | Method of treating diabetic nephropathy by administering antibodies to vascular endothelial growth factor B (VEGF-B) |
CA2931978A1 (en) | 2013-12-02 | 2015-06-11 | Abbvie Inc. | Compositions and methods for treating osteoarthritis |
US9309314B2 (en) | 2013-12-03 | 2016-04-12 | Agency For Science, Technology And Research (A*Star) | Polypeptides, nucleic acids and uses thereof |
EP3611191A1 (en) | 2013-12-09 | 2020-02-19 | Allakos Inc. | Anti-siglec-8 antibodies and methods of use thereof |
SG11201604784XA (en) | 2013-12-13 | 2016-07-28 | Genentech Inc | Anti-cd33 antibodies and immunoconjugates |
EP2883883A1 (en) | 2013-12-16 | 2015-06-17 | Cardio3 Biosciences S.A. | Therapeutic targets and agents useful in treating ischemia reperfusion injury |
CN105899535A (en) | 2013-12-17 | 2016-08-24 | 豪夫迈·罗氏有限公司 | Methods of treating cancer using pd-1 axis binding antagonists and an anti-cd20 antibody |
PE20210648A1 (en) | 2013-12-17 | 2021-03-26 | 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. |
CA2934028A1 (en) | 2013-12-17 | 2015-06-25 | Genentech, Inc. | Combination therapy comprising ox40 binding agonists and pd-1 axis binding antagonists |
EP3082860B1 (en) | 2013-12-18 | 2020-11-25 | CSL Limited | Method of treating wounds |
MX2016007208A (en) | 2013-12-20 | 2016-07-21 | Hoffmann La Roche | HUMANIZED ANTI-Tau(pS422) ANTIBODIES AND METHODS OF USE. |
TWI670283B (en) | 2013-12-23 | 2019-09-01 | 美商建南德克公司 | Antibodies and methods of use |
BR112016014945A2 (en) | 2014-01-03 | 2018-01-23 | F. Hoffmann-La Roche Ag | conjugate, pharmaceutical formulation and use |
EP3089758B1 (en) | 2014-01-03 | 2021-01-27 | F.Hoffmann-La Roche Ag | Covalently linked helicar-anti-helicar antibody conjugates and uses thereof |
WO2015101586A1 (en) | 2014-01-03 | 2015-07-09 | F. Hoffmann-La Roche Ag | Bispecific anti-hapten/anti-blood brain barrier receptor antibodies, complexes thereof and their use as blood brain barrier shuttles |
JP6557664B2 (en) | 2014-01-06 | 2019-08-07 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Monovalent blood-brain barrier shuttle module |
SG11201605455YA (en) | 2014-01-10 | 2016-08-30 | Birdie Biopharmaceuticals Inc | Compounds and compositions for treating egfr expressing tumors |
WO2015112909A1 (en) | 2014-01-24 | 2015-07-30 | Genentech, Inc. | Methods of using anti-steap1 antibodies and immunoconjugates |
AU2015214264B2 (en) | 2014-02-04 | 2018-12-20 | Curis, Inc. | Mutant Smoothened and methods of using the same |
JP6685912B2 (en) | 2014-02-08 | 2020-04-22 | ジェネンテック, インコーポレイテッド | Alzheimer's disease treatment method |
EP3102230B1 (en) | 2014-02-08 | 2021-04-28 | F. Hoffmann-La Roche AG | Methods of treating alzheimer's disease |
AU2015217271B2 (en) | 2014-02-12 | 2018-10-25 | Genentech, Inc. | Anti-Jagged1 antibodies and methods of use |
RU2019119857A (en) | 2014-02-20 | 2019-07-12 | Аллерган, Инк. | ANTIBODY TO C5 COMPLETE COMPONENT |
MX2016010729A (en) | 2014-02-21 | 2016-10-26 | Genentech Inc | Anti-il-13/il-17 bispecific antibodies and uses thereof. |
AU2015223056B2 (en) | 2014-02-27 | 2020-10-01 | Allergan, Inc. | Complement Factor Bb antibodies |
DK3110446T3 (en) | 2014-02-28 | 2022-02-28 | Allakos Inc | Methods and compositions for treating Siglec-8-associated diseases |
JP6644717B2 (en) | 2014-03-14 | 2020-02-12 | ジェネンテック, インコーポレイテッド | Methods and compositions for secreting heterologous polypeptides |
BR112016021717A2 (en) | 2014-03-21 | 2018-07-10 | Abbvie Inc | anti-egfr antibodies and antibody-drug conjugates |
WO2015140591A1 (en) | 2014-03-21 | 2015-09-24 | Nordlandssykehuset Hf | Anti-cd14 antibodies and uses thereof |
MX2016012285A (en) | 2014-03-24 | 2017-01-23 | Genentech Inc | Cancer treatment with c-met antagonists and correlation of the latter with hgf expression. |
CN106132439A (en) | 2014-03-31 | 2016-11-16 | 豪夫迈·罗氏有限公司 | Comprise antiangiogenic agent and OX40 combines the combination treatment of agonist |
MA40682B1 (en) | 2014-03-31 | 2020-01-31 | Hoffmann La Roche | Anti-ox40 antibodies and methods of use thereof |
AP2016009549A0 (en) | 2014-04-18 | 2016-11-30 | Acceleron Pharma Inc | Methods for increasing red blood cell levels and treating sickle-cell disease |
WO2015164615A1 (en) | 2014-04-24 | 2015-10-29 | University Of Oslo | Anti-gluten antibodies and uses thereof |
UA119352C2 (en) | 2014-05-01 | 2019-06-10 | Тева Фармасьютикалз Острейліа Пті Лтд | Combination of lenalidomide or pomalidomide and cd38 antibody-attenuated interferon-alpha constructs, and the use thereof |
CN106415269B (en) | 2014-05-08 | 2020-11-27 | 贵州美鑫达医疗科技有限公司 | Direct immunohistochemical assay |
CN106413750B (en) | 2014-05-16 | 2022-04-29 | 免疫医疗有限责任公司 | Molecules with altered neonatal Fc receptor binding with enhanced therapeutic and diagnostic properties |
WO2015179658A2 (en) | 2014-05-22 | 2015-11-26 | Genentech, Inc. | Anti-gpc3 antibodies and immunoconjugates |
KR20170005016A (en) | 2014-05-23 | 2017-01-11 | 제넨테크, 인크. | Mit biomarkers and methods using the same |
US11033620B2 (en) | 2014-06-09 | 2021-06-15 | Biomed Valley Discoveries, Inc. | Combination therapies targeting tumor-associated stroma or tumor cells and microtubules |
WO2015191583A2 (en) | 2014-06-09 | 2015-12-17 | Biomed Valley Discoveries, Inc. | Combination therapies targeting tumor-associated stroma or tumor cells and topoisomerase |
US10758613B2 (en) | 2014-06-09 | 2020-09-01 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services National Intstitutes Of Health | Combination therapies using anti-metabolites and agents that target tumor-associated stroma or tumor cells |
US10758526B2 (en) | 2014-06-09 | 2020-09-01 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services National Institutes Of Health | Combination therapies using agents that target tumor-associated stroma or tumor cells and other pathways |
WO2015191596A1 (en) | 2014-06-09 | 2015-12-17 | Biomed Valley Discoveries, Inc. | Combination therapies using platinum agents and agents that target tumor-associated stroma or tumor cells |
US11034757B2 (en) | 2014-06-09 | 2021-06-15 | Biomed Valley Discoveries, Inc. | Combination therapies using agents that target tumor-associated stroma or tumor cells and tumor vasculature |
WO2015191615A2 (en) | 2014-06-09 | 2015-12-17 | Biomed Valley Discoveries, Inc. | Combination therapies using agents that target tumor-associated stroma or tumor cells and alkylating agents |
CN106459202A (en) | 2014-06-11 | 2017-02-22 | 豪夫迈·罗氏有限公司 | Anti-lgR5 antibodies and uses thereof |
WO2015191986A1 (en) | 2014-06-13 | 2015-12-17 | Genentech, Inc. | Methods of treating and preventing cancer drug resistance |
BR122023023170A2 (en) | 2014-06-13 | 2024-02-20 | Acceleron Pharma Inc. | USE OF AN ACTRII ANTAGONIST IN THE TREATMENT OR PREVENTION OF SKIN ULCERS ASSOCIATED WITH BETA-THALASSEMIA |
NL2013007B1 (en) | 2014-06-16 | 2016-07-05 | Ablynx Nv | Methods of treating TTP with immunoglobulin single variable domains and uses thereof. |
NL2013661B1 (en) | 2014-10-21 | 2016-10-05 | Ablynx Nv | KV1.3 Binding immunoglobulins. |
AR100978A1 (en) | 2014-06-26 | 2016-11-16 | Hoffmann La Roche | ANTI-Tau HUMANIZED ANTIBODY BRAIN LAUNCHERS (pS422) AND USES OF THE SAME |
MX2016015280A (en) | 2014-06-26 | 2017-03-03 | Hoffmann La Roche | Anti-brdu antibodies and methods of use. |
JP6694400B2 (en) | 2014-07-03 | 2020-05-13 | ジェネンテック, インコーポレイテッド | Polypeptide expression system |
AU2015286043B2 (en) | 2014-07-09 | 2020-08-20 | Birdie Biopharmaceuticals Inc. | Anti-PD-L1 combinations for treating tumors |
CN105233291A (en) | 2014-07-09 | 2016-01-13 | 博笛生物科技有限公司 | Combined therapy composition and combined therapy method for treating cancers |
CN105440135A (en) | 2014-09-01 | 2016-03-30 | 博笛生物科技有限公司 | Anti-PD-L1 conjugate for treating tumors |
SG11201700207WA (en) | 2014-07-11 | 2017-02-27 | Genentech Inc | Anti-pd-l1 antibodies and diagnostic uses thereof |
EP3166627A1 (en) | 2014-07-11 | 2017-05-17 | Genentech, Inc. | Notch pathway inhibition |
JP6671363B2 (en) | 2014-07-15 | 2020-03-25 | イッサム リサーチ ディヴェロップメント カンパニー オブ ザ ヘブリュー ユニバーシティー オブ エルサレム リミティッド | Isolated polypeptide of CD44 and uses thereof |
KR102317315B1 (en) | 2014-08-04 | 2021-10-27 | 에프. 호프만-라 로슈 아게 | Bispecific t cell activating antigen binding molecules |
AU2015308818B2 (en) | 2014-08-28 | 2021-02-25 | Bioatla Llc | Conditionally active chimeric antigen receptors for modified T-cells |
DK3191135T3 (en) | 2014-09-12 | 2020-10-12 | Genentech Inc | Anti-HER2 antibodies and immunoconjugates |
EP3191518B1 (en) | 2014-09-12 | 2020-01-15 | Genentech, Inc. | Anti-b7-h4 antibodies and immunoconjugates |
EP3693391A1 (en) | 2014-09-12 | 2020-08-12 | Genentech, Inc. | Anti-cll-1 antibodies and immunoconjugates |
EP3193932B1 (en) | 2014-09-15 | 2023-04-26 | F. Hoffmann-La Roche AG | Antibody formulations |
CN107124870A (en) | 2014-09-17 | 2017-09-01 | 基因泰克公司 | Immunoconjugates comprising Anti-HER 2 and Pyrrolobenzodiazepines * |
US20160082120A1 (en) | 2014-09-23 | 2016-03-24 | Genentech, Inc. | METHODS OF USING ANTI-CD79b IMMUNOCONJUGATES |
RU2700650C2 (en) | 2014-10-10 | 2019-09-18 | Аблинкс Н.В. | Inhaler device for use in aerosol therapy of respiratory diseases |
RU2017115670A (en) | 2014-10-10 | 2018-11-15 | Аблинкс Н.В. | RSV INFECTION TREATMENT |
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 |
EP3209697A4 (en) | 2014-10-23 | 2018-05-30 | La Trobe University | Fn14-binding proteins and uses thereof |
AU2015336931B2 (en) | 2014-10-23 | 2021-04-29 | Kira Biotech Pty Limited | CD83 binding proteins and uses thereof |
WO2016065409A1 (en) | 2014-10-29 | 2016-05-06 | Teva Pharmaceuticals Australia Pty Ltd | INTERFERON α2B VARIANTS |
KR20170072884A (en) | 2014-10-29 | 2017-06-27 | 시애틀 지네틱스, 인크. | Dosage and administration of non-fucosylated anti-cd40 antibodies |
WO2016073378A1 (en) | 2014-11-03 | 2016-05-12 | Genentech, Inc. | Assays for detecting t cell immune subsets and methods of use thereof |
MX2017005751A (en) | 2014-11-03 | 2018-04-10 | Genentech Inc | Method and biomarkers for predicting efficacy and evaluation of an ox40 agonist treatment. |
KR102544705B1 (en) | 2014-11-05 | 2023-06-15 | 제넨테크, 인크. | Methods of producing two chain proteins in bacteria |
BR112017009330A2 (en) | 2014-11-05 | 2017-12-19 | Agrosavfe N V | transgenic plant comprising a polynucleotide encoding a heavy chain antibody variable domain |
CN108064308B (en) | 2014-11-05 | 2023-06-09 | 豪夫迈·罗氏有限公司 | Method for producing double-stranded protein in bacteria |
WO2016073157A1 (en) | 2014-11-06 | 2016-05-12 | Genentech, Inc. | Anti-ang2 antibodies and methods of use thereof |
AU2015343494A1 (en) | 2014-11-06 | 2017-04-27 | Genentech, Inc. | Combination therapy comprising OX40 binding agonists and TIGIT inhibitors |
EP3217787B1 (en) | 2014-11-10 | 2019-04-17 | F.Hoffmann-La Roche Ag | Animal model for nephropathy and agents for treating the same |
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 |
CN113372434A (en) | 2014-11-14 | 2021-09-10 | 豪夫迈·罗氏有限公司 | Antigen binding molecules comprising TNF family ligand trimers |
CN107429075B (en) | 2014-11-17 | 2022-11-01 | 卡内基梅隆大学 | Activatable two-component photosensitizer |
WO2016081384A1 (en) | 2014-11-17 | 2016-05-26 | Genentech, Inc. | Combination therapy comprising ox40 binding agonists and pd-1 axis binding antagonists |
CN107250158B (en) | 2014-11-19 | 2022-03-25 | 基因泰克公司 | Anti-transferrin receptor/anti-BACE 1 multispecific antibodies and methods of use |
CN107108745B (en) | 2014-11-19 | 2021-01-12 | 基因泰克公司 | Antibodies against BACE1 and their use for immunotherapy of neurological diseases |
EP3221362B1 (en) | 2014-11-19 | 2019-07-24 | F.Hoffmann-La Roche Ag | Anti-transferrin receptor antibodies and methods of use |
WO2016079081A1 (en) | 2014-11-20 | 2016-05-26 | F. Hoffmann-La Roche Ag | Common light chains and methods of use |
KR20240024318A (en) | 2014-11-20 | 2024-02-23 | 에프. 호프만-라 로슈 아게 | Combination therapy of t cell activating bispecific antigen binding molecules cd3 abd folate receptor 1 (folr1) and pd-1 axis binding antagonists |
MA41119A (en) | 2014-12-03 | 2017-10-10 | Acceleron Pharma Inc | METHODS OF TREATMENT OF MYELODYSPLASIC SYNDROMES AND SIDEROBLASTIC ANEMIA |
ES2764111T3 (en) | 2014-12-03 | 2020-06-02 | Hoffmann La Roche | Multispecific antibodies |
ES2744540T3 (en) | 2014-12-05 | 2020-02-25 | Hoffmann La Roche | Anti-CD79b antibodies and usage procedures |
BR112017011234A2 (en) | 2014-12-10 | 2018-03-27 | Genentech Inc | antibodies to the blood-brain barrier receptor and methods of use |
WO2016094881A2 (en) | 2014-12-11 | 2016-06-16 | Abbvie Inc. | Lrp-8 binding proteins |
CA2971278C (en) | 2014-12-19 | 2023-09-19 | Ablynx N.V. | Cysteine linked nanobody dimers |
AU2015367224B2 (en) | 2014-12-19 | 2020-12-10 | Monash University | IL-21 antibodies |
US9765135B2 (en) | 2014-12-19 | 2017-09-19 | Chugai Seiyaku Kabushiki Kaisha | Anti-C5 antibodies |
US10435467B2 (en) | 2015-01-08 | 2019-10-08 | Biogen Ma Inc. | LINGO-1 antagonists and uses for treatment of demyelinating disorders |
SG10202111844VA (en) | 2015-01-09 | 2021-12-30 | Adalta Ltd | Cxcr4 binding molecules |
CN113956354A (en) | 2015-01-22 | 2022-01-21 | 中外制药株式会社 | Combinations and methods of use of two or more anti-C5 antibodies |
WO2016118947A1 (en) | 2015-01-23 | 2016-07-28 | The University Of North Carolina At Chapel Hill | Apparatuses, systems, and methods for preclinical ultrasound imaging of subjects |
CN107531780B (en) | 2015-02-03 | 2021-11-02 | 国家健康与医学研究院 | Conformational single domain antibodies to Rho GTPase and uses thereof |
JP2018512597A (en) | 2015-02-04 | 2018-05-17 | ジェネンテック, インコーポレイテッド | Mutant smoothened and method of using the same |
WO2016125495A1 (en) | 2015-02-05 | 2016-08-11 | Chugai Seiyaku Kabushiki Kaisha | Antibodies comprising an ion concentration dependent antigen-binding domain, fc region variants, il-8-binding antibodies, and uses therof |
AU2016219511B2 (en) | 2015-02-09 | 2020-11-12 | Research Development Foundation | Engineered immunoglobulin Fc polypeptides displaying improved complement activation |
US20170151281A1 (en) | 2015-02-19 | 2017-06-01 | Batu Biologics, Inc. | Chimeric antigen receptor dendritic cell (car-dc) for treatment of cancer |
WO2016135041A1 (en) | 2015-02-26 | 2016-09-01 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Fusion proteins and antibodies comprising thereof for promoting apoptosis |
AR103935A1 (en) | 2015-03-16 | 2017-06-14 | Genentech Inc | METHODS OF DETECTION AND QUANTIFICATION OF IL-13 AND ITS USES IN THE DIAGNOSIS AND TREATMENT OF DISEASES ASSOCIATED WITH TH-2 |
WO2016146833A1 (en) | 2015-03-19 | 2016-09-22 | F. Hoffmann-La Roche Ag | Biomarkers for nad(+)-diphthamide adp ribosyltransferase resistance |
MA41919A (en) | 2015-04-06 | 2018-02-13 | Acceleron Pharma Inc | ALK4 HETEROMULTIMERS: ACTRIIB AND THEIR USES |
MA53400A (en) | 2015-04-06 | 2021-08-04 | Acceleron Pharma Inc | ALK7 HETEROMULTIMERS: ACTRIIB AND THEIR USES |
CA2981183A1 (en) | 2015-04-07 | 2016-10-13 | Greg Lazar | Antigen binding complex having agonistic activity and methods of use |
SI3280441T1 (en) | 2015-04-07 | 2022-01-31 | Alector Llc | Anti-sortilin antibodies and methods of use thereof |
WO2016172160A1 (en) | 2015-04-21 | 2016-10-27 | Genentech, Inc. | Compositions and methods for prostate cancer analysis |
WO2016172551A2 (en) | 2015-04-24 | 2016-10-27 | Genentech, Inc. | Methods of identifying bacteria comprising binding polypeptides |
KR20170138570A (en) | 2015-04-30 | 2017-12-15 | 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 | Anti-aP2 antibodies and antigen binding substances for the treatment of metabolic disorders |
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 |
EP3936524A3 (en) | 2015-05-11 | 2022-06-15 | F. Hoffmann-La Roche AG | Compositions and methods of treating lupus nephritis |
AU2016262074A1 (en) | 2015-05-12 | 2017-11-09 | Genentech, Inc. | Therapeutic and diagnostic methods for cancer |
EP3095465A1 (en) | 2015-05-19 | 2016-11-23 | U3 Pharma GmbH | Combination of fgfr4-inhibitor and bile acid sequestrant |
PE20180193A1 (en) | 2015-05-29 | 2018-01-26 | Abbvie Inc | ANTI-CD40 ANTIBODIES AND THEIR USES |
ES2789500T5 (en) | 2015-05-29 | 2023-09-20 | Hoffmann La Roche | Therapeutic and diagnostic procedures for 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 |
KR20180011117A (en) | 2015-05-31 | 2018-01-31 | 큐어제닉스 코포레이션 | Composite composition for immunotherapy |
WO2016196679A1 (en) | 2015-06-02 | 2016-12-08 | Genentech, Inc. | Compositions and methods for using anti-il-34 antibodies to treat neurological diseases |
SG10201911349YA (en) | 2015-06-05 | 2020-01-30 | Genentech Inc | Anti-tau antibodies and methods of use |
CN107810011A (en) | 2015-06-08 | 2018-03-16 | 豪夫迈·罗氏有限公司 | Use the method for anti-OX40 antibodies for treating cancer |
US20170000885A1 (en) | 2015-06-08 | 2017-01-05 | Genentech, Inc. | Methods of treating cancer using anti-ox40 antibodies and pd-1 axis binding antagonists |
AU2016276981B2 (en) | 2015-06-12 | 2022-10-06 | Alector Llc | Anti-CD33 antibodies and methods of use thereof |
JP2018518491A (en) | 2015-06-12 | 2018-07-12 | アレクトル エルエルシー | Anti-CD33 antibody and method of use thereof |
EP3307780A1 (en) | 2015-06-15 | 2018-04-18 | Genentech, Inc. | Antibodies and immunoconjugates |
TW201710286A (en) | 2015-06-15 | 2017-03-16 | 艾伯維有限公司 | Binding proteins against VEGF, PDGF, and/or their receptors |
EP3310811B1 (en) | 2015-06-16 | 2021-06-16 | Genentech, Inc. | Anti-cd3 antibodies and methods of use |
CN107847568B (en) | 2015-06-16 | 2022-12-20 | 豪夫迈·罗氏有限公司 | anti-CLL-1 antibodies and methods of use |
SI3310814T1 (en) | 2015-06-16 | 2023-11-30 | F. Hoffmann - La Roche Ag | Humanized and affinity matured antibodies to fcrh5 and methods of use |
EP3310812A2 (en) | 2015-06-17 | 2018-04-25 | H. Hoffnabb-La Roche Ag | Anti-her2 antibodies and methods of use |
JP6896650B2 (en) | 2015-06-17 | 2021-06-30 | ジェネンテック, インコーポレイテッド | Treatment of Locally Advanced or Metastatic Breast Cancer Using PD-1 Axle Antagonists and Taxanes |
EP3310385A4 (en) | 2015-06-17 | 2018-12-19 | Allakos Inc. | Methods and compositions for treating fibrotic diseases |
CN107810196B (en) | 2015-06-24 | 2021-11-05 | 豪夫迈·罗氏有限公司 | Humanized anti-Tau (pS422) antibodies and methods of use |
CN108473573A (en) | 2015-06-29 | 2018-08-31 | 豪夫迈·罗氏有限公司 | II type anti-CD 20 antibodies are used in organ transplant |
JP6786529B2 (en) | 2015-06-29 | 2020-11-18 | バイオメッド バレー ディスカバリーズ,インコーポレイティド | Combination of LPT-723 and Immune Checkpoint Inhibitors and Methods of Treatment |
EP3514174B1 (en) | 2015-06-29 | 2021-03-31 | Ventana Medical Systems, Inc. | Materials and methods for performing histochemical assays for human pro-epiregulin and amphiregulin |
WO2017004330A1 (en) | 2015-06-30 | 2017-01-05 | Seattle Genetics, Inc. | Anti-ntb-a antibodies and related compositions and methods |
WO2017023866A1 (en) | 2015-07-31 | 2017-02-09 | Boston Biomedical, Inc. | Method of targeting stat3 and other non-druggable proteins |
TW202330904A (en) | 2015-08-04 | 2023-08-01 | 美商再生元醫藥公司 | Taurine supplemented cell culture medium and methods of use |
AU2016301380B2 (en) | 2015-08-04 | 2021-07-01 | Acceleron Pharma Inc. | Methods for treating myeloproliferative disorders |
CA2994951A1 (en) | 2015-08-07 | 2017-02-16 | 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 |
MX2018003005A (en) | 2015-09-18 | 2018-04-11 | Chugai Pharmaceutical Co Ltd | Il-8-binding antibodies and uses thereof. |
US11078251B2 (en) | 2015-09-18 | 2021-08-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | T cell receptors (TCR) and uses thereof for the diagnosis and treatment of diabetes |
CA2999369C (en) | 2015-09-22 | 2023-11-07 | Spring Bioscience Corporation | Anti-ox40 antibodies and diagnostic uses thereof |
SG10201911226QA (en) | 2015-09-23 | 2020-01-30 | Genentech Inc | Optimized variants of anti-vegf antibodies |
ES2768957T3 (en) | 2015-09-24 | 2020-06-24 | Abvitro Llc | HIV Antibody Compositions and Methods of Use |
US10947598B2 (en) | 2015-09-29 | 2021-03-16 | Inserm (Institut National De La Sante Et De La Recherche Medicale) | Methods for determining the metabolic status of lymphomas |
AR106188A1 (en) | 2015-10-01 | 2017-12-20 | Hoffmann La Roche | ANTI-CD19 HUMANIZED HUMAN ANTIBODIES AND METHODS OF USE |
WO2017055385A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Anti-cd3xgd2 bispecific t cell activating antigen binding molecules |
WO2017055392A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Anti-cd3xcd44v6 bispecific t cell activating antigen binding molecules |
WO2017055393A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Anti-cd3xtim-3 bispecific t cell activating antigen binding molecules |
MA43345A (en) | 2015-10-02 | 2018-08-08 | Hoffmann La Roche | PYRROLOBENZODIAZEPINE ANTIBODY-DRUG CONJUGATES AND METHODS OF USE |
WO2017055314A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Bispecific anti-cd19xcd3 t cell activating antigen binding molecules |
US20170096485A1 (en) | 2015-10-02 | 2017-04-06 | Hoffmann-La Roche Inc. | Bispecific t cell activating antigen binding molecules |
EP3356410B1 (en) | 2015-10-02 | 2021-10-20 | F. Hoffmann-La Roche AG | Bispecific anti-ceaxcd3 t cell activating antigen binding molecules |
KR102146319B1 (en) | 2015-10-02 | 2020-08-25 | 에프. 호프만-라 로슈 아게 | Bispecific antibodies specific for PD1 and TIM3 |
WO2017055395A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Anti-cd3xrob04 bispecific t cell activating antigen binding molecules |
UA123826C2 (en) | 2015-10-02 | 2021-06-09 | Ф. Хоффманн-Ля Рош Аг | Anti-pd1 antibodies and methods of use |
CN107949574A (en) | 2015-10-02 | 2018-04-20 | 豪夫迈·罗氏有限公司 | Bispecific T cell activation antigen binding molecules |
EP3356403A2 (en) | 2015-10-02 | 2018-08-08 | H. Hoffnabb-La Roche Ag | Bispecific antibodies specific for a costimulatory tnf receptor |
MX2018004157A (en) | 2015-10-07 | 2019-04-01 | F Hoffmann La Roche Ag | Bispecific antibodies with tetravalency for a costimulatory tnf receptor. |
US10556953B2 (en) | 2015-10-12 | 2020-02-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Agent capable of depleting CD8 T cells for the treatment of myocardial infarction or acute myocardial infarction |
MA43354A (en) | 2015-10-16 | 2018-08-22 | Genentech Inc | CONJUGATE DRUG CONJUGATES WITH CLOUDY DISULPHIDE |
MA45326A (en) | 2015-10-20 | 2018-08-29 | Genentech Inc | CALICHEAMICIN-ANTIBODY-DRUG CONJUGATES AND METHODS OF USE |
US10604577B2 (en) | 2015-10-22 | 2020-03-31 | Allakos Inc. | Methods and compositions for treating systemic mastocytosis |
EP3184547A1 (en) | 2015-10-29 | 2017-06-28 | F. Hoffmann-La Roche AG | Anti-tpbg antibodies and methods of use |
AU2016344665C1 (en) | 2015-10-29 | 2023-07-27 | F. Hoffmann-La Roche Ag | Anti-variant Fc-region antibodies and methods of use |
US10407510B2 (en) | 2015-10-30 | 2019-09-10 | Genentech, Inc. | Anti-factor D antibodies and conjugates |
KR102162324B1 (en) | 2015-10-30 | 2020-10-07 | 제넨테크, 인크. | Anti-HtrA1 antibodies and methods of use thereof |
EP3371217A1 (en) | 2015-11-08 | 2018-09-12 | H. Hoffnabb-La Roche Ag | Methods of screening for multispecific antibodies |
EP3374389A1 (en) | 2015-11-13 | 2018-09-19 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Anti- nkg2d single domain antibodies and uses thereof |
KR20180083402A (en) | 2015-11-20 | 2018-07-20 | 오스트레일리언 바이오메디컬 컴퍼니 피티와이 리미티드 | Compounds for medical use |
EP3380121B1 (en) | 2015-11-23 | 2023-12-20 | Acceleron Pharma Inc. | Actrii antagonist for use in treating eye disorders |
CA3006398A1 (en) | 2015-11-27 | 2017-06-01 | Ablynx Nv | Polypeptides inhibiting cd40l |
KR20180085740A (en) | 2015-12-09 | 2018-07-27 | 에프. 호프만-라 로슈 아게 | Type II anti-CD20 antibodies to reduce the formation of anti-drug antibodies |
EP3178848A1 (en) | 2015-12-09 | 2017-06-14 | F. Hoffmann-La Roche AG | Type ii anti-cd20 antibody for reducing formation of anti-drug antibodies |
AU2016372930B2 (en) | 2015-12-18 | 2020-10-08 | Chugai Seiyaku Kabushiki Kaisha | Anti-C5 antibodies and methods of use |
CN108602878B (en) | 2015-12-18 | 2022-06-28 | 卫材R&D管理有限公司 | C-terminal lysine conjugated immunoglobulins |
JP7008023B2 (en) | 2015-12-30 | 2022-01-25 | ジェネンテック, インコーポレイテッド | Formulation with reduced polysorbate degradation |
MX2018008063A (en) | 2015-12-30 | 2018-11-29 | Genentech Inc | Use of tryptophan derivatives for protein formulations. |
WO2017118634A1 (en) | 2016-01-04 | 2017-07-13 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of pd-1 and tim-3 as a measure for cd8+ cells in predicting and treating renal cell carcinoma |
CN107531795B (en) | 2016-01-05 | 2021-01-19 | 江苏恒瑞医药股份有限公司 | PCSK9 antibody, antigen-binding fragment thereof and medical application thereof |
US10596257B2 (en) | 2016-01-08 | 2020-03-24 | Hoffmann-La Roche Inc. | Methods of treating CEA-positive cancers using PD-1 axis binding antagonists and anti-CEA/anti-CD3 bispecific antibodies |
WO2017127764A1 (en) | 2016-01-20 | 2017-07-27 | Genentech, Inc. | High dose treatments for alzheimer's disease |
WO2017129558A1 (en) | 2016-01-25 | 2017-08-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for predicting or treating myelopoiesis-driven cardiometabolic diseases and sepsis |
US10822415B2 (en) | 2016-01-28 | 2020-11-03 | Inserm (Institut National De La Santéet De La Recherche Médicale) | Methods for enhancing the potency of the immune checkpoint inhibitors |
ES2924775T3 (en) | 2016-01-28 | 2022-10-10 | Inst Nat Sante Rech Med | Methods and pharmaceutical composition for the treatment of cancer |
WO2017129763A1 (en) | 2016-01-28 | 2017-08-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for the treatment of signet ring cell gastric cancer |
EP3411396A1 (en) | 2016-02-04 | 2018-12-12 | Curis, Inc. | Mutant smoothened and methods of using the same |
CN109196121B (en) | 2016-02-29 | 2022-01-04 | 基因泰克公司 | Methods for treatment and diagnosis of cancer |
KR102445255B1 (en) | 2016-03-02 | 2022-09-22 | 에자이 알앤드디 매니지먼트 가부시키가이샤 | Eribulin-Based Antibody-Drug Conjugates and Methods of Use |
CN116196412A (en) | 2016-03-15 | 2023-06-02 | 中外制药株式会社 | Methods of treating cancer using PD-1 axis binding antagonists and anti-GPC 3 antibodies |
WO2017160954A1 (en) | 2016-03-15 | 2017-09-21 | Seattle Genetics, Inc. | Combinations of pbd-based antibody drug conjugates with bcl-2 inhibitors |
CA3012422A1 (en) | 2016-03-22 | 2017-09-28 | F. Hoffmann-La Roche Ag | Protease-activated t cell bispecific molecules |
PE20231511A1 (en) | 2016-03-22 | 2023-09-26 | Hoffmann La Roche | BISPECIFIC T CELL MOLECULES ACTIVATED BY PROTEASES THAT BIND SPECIFICALLY TO FOLATE RECEPTOR 1 (FOLR1) AND CD3 |
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 |
EP3436477A2 (en) | 2016-03-29 | 2019-02-06 | Janssen Biotech, Inc. | Method of treating psoriasis with increased interval dosing of anti-il12 and/or -23 antibody |
EP3231813A1 (en) | 2016-03-29 | 2017-10-18 | F. Hoffmann-La Roche AG | Trimeric costimulatory tnf family ligand-containing antigen binding molecules |
EP3439659A1 (en) | 2016-04-06 | 2019-02-13 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for the treatment of age-related cardiometabolic diseases |
EP3443004A1 (en) | 2016-04-14 | 2019-02-20 | H. Hoffnabb-La Roche Ag | Anti-rspo3 antibodies and methods of use |
PL3443350T3 (en) | 2016-04-15 | 2021-05-31 | F. Hoffmann-La Roche Ag | Methods for monitoring and treating cancer |
KR20190003957A (en) | 2016-04-15 | 2019-01-10 | 제넨테크, 인크. | Cancer monitoring and treatment methods |
WO2017182609A1 (en) | 2016-04-22 | 2017-10-26 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical composition for the treatment of inflammatory skin diseases associated with desmoglein-1 deficiency |
CN114617962A (en) | 2016-04-27 | 2022-06-14 | 艾伯维公司 | Methods of using anti-IL-13 antibodies to treat diseases in which IL-13 activity is detrimental |
CR20180509A (en) | 2016-05-02 | 2019-02-15 | Hoffmann La Roche | CONTORSBODY - A BIND OF DIANA MONOCATENARY |
CN109311968A (en) | 2016-05-02 | 2019-02-05 | 埃博灵克斯股份有限公司 | Treat rsv infection |
US11098124B2 (en) | 2016-05-03 | 2021-08-24 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | CD31 shed as a molecular target for imaging of inflammation |
EP3744348B1 (en) | 2016-05-06 | 2022-03-23 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Pharmaceutical compositions for the treatment of chemoresistant acute myeloid leukemia (aml) |
WO2017194554A1 (en) | 2016-05-10 | 2017-11-16 | Inserm (Institut National De La Sante Et De La Recherche Medicale) | Combinations therapies for the treatment of cancer |
WO2017194441A1 (en) | 2016-05-11 | 2017-11-16 | F. Hoffmann-La Roche Ag | Modified anti-tenascin antibodies and methods of use |
WO2017194442A1 (en) | 2016-05-11 | 2017-11-16 | F. Hoffmann-La Roche Ag | Antigen binding molecules comprising a tnf family ligand trimer and a tenascin binding moiety |
EP3243836A1 (en) | 2016-05-11 | 2017-11-15 | F. Hoffmann-La Roche AG | C-terminally fused tnf family ligand trimer-containing antigen binding molecules |
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 |
EP4122958A1 (en) | 2016-05-13 | 2023-01-25 | BioAtla, Inc. | Anti-ror2 antibodies, antibody fragments, their immunoconjugates and uses thereof |
BR112018072986A2 (en) | 2016-05-18 | 2019-03-06 | Boehringer Ingelheim International Gmbh | anti-pd-1 and anti-lag3 antibodies for cancer treatment |
ES2858151T3 (en) | 2016-05-20 | 2021-09-29 | Hoffmann La Roche | PROTAC-Antibody Conjugates and Procedures for Use |
WO2017202962A1 (en) | 2016-05-24 | 2017-11-30 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for the treatment of non small cell lung cancer (nsclc) that coexists with chronic obstructive pulmonary disease (copd) |
WO2017205741A1 (en) | 2016-05-27 | 2017-11-30 | Genentech, Inc. | Bioanalytical method for the characterization of site-specific antibody-drug conjugates |
WO2017202890A1 (en) | 2016-05-27 | 2017-11-30 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for predicting and treating myeloma |
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 |
CA3059010A1 (en) | 2016-06-02 | 2018-12-06 | F. Hoffmann-La Roche Ag | Type ii anti-cd20 antibody and anti-cd20/cd3 bispecific antibody for treatment of cancer |
WO2017214024A1 (en) | 2016-06-06 | 2017-12-14 | Genentech, Inc. | Silvestrol antibody-drug conjugates and methods of use |
CN109562168A (en) | 2016-06-08 | 2019-04-02 | 艾伯维公司 | Anti-CD 98 antibody and antibody drug conjugates |
PL3458479T3 (en) | 2016-06-08 | 2021-07-26 | Abbvie Inc. | Anti-b7-h3 antibodies and antibody drug conjugates |
WO2017214301A1 (en) | 2016-06-08 | 2017-12-14 | Abbvie Inc. | Anti-egfr antibody drug conjugates |
AU2017279539A1 (en) | 2016-06-08 | 2019-01-03 | Abbvie Inc. | Anti-B7-H3 antibodies and antibody drug conjugates |
BR112018075639A2 (en) | 2016-06-08 | 2019-04-09 | Abbvie Inc. | anti-egfr drug antibody conjugates |
JP2019521973A (en) | 2016-06-08 | 2019-08-08 | アッヴィ・インコーポレイテッド | Anti-BH7-H3 antibody and antibody drug conjugate |
JP2019524651A (en) | 2016-06-08 | 2019-09-05 | アッヴィ・インコーポレイテッド | Anti-CD98 antibodies and antibody drug conjugates |
CN109562170B (en) | 2016-06-08 | 2023-01-13 | 艾伯维公司 | anti-CD 98 antibodies and antibody drug conjugates |
JP2019521106A (en) | 2016-06-08 | 2019-07-25 | アッヴィ・インコーポレイテッド | Anti-EGFR antibody drug conjugate |
GB201610198D0 (en) | 2016-06-10 | 2016-07-27 | Ucb Biopharma Sprl | Anti-ige antibodies |
MX2018015331A (en) | 2016-06-10 | 2019-08-16 | Eisai R&D Man Co Ltd | Lysine conjugated immunoglobulins. |
CN116143918A (en) | 2016-06-24 | 2023-05-23 | 豪夫迈·罗氏有限公司 | Anti-polyubiquitin multispecific antibodies |
WO2018007442A1 (en) | 2016-07-06 | 2018-01-11 | Ablynx N.V. | Treatment of il-6r related diseases |
WO2018013936A1 (en) | 2016-07-15 | 2018-01-18 | Acceleron Pharma Inc. | Compositions and methods for treating pulmonary hypertension |
EP3487522A4 (en) | 2016-07-19 | 2020-04-01 | Teva Pharmaceuticals Australia Pty Ltd | Anti-cd47 combination therapy |
WO2018014260A1 (en) | 2016-07-20 | 2018-01-25 | Nanjing Legend Biotech Co., Ltd. | Multispecific antigen binding proteins and methods of use thereof |
BR112019001615A2 (en) | 2016-07-27 | 2019-04-30 | Acceleron Pharma Inc. | methods and compositions for treating myelofibrosis |
US20190185578A1 (en) | 2016-07-29 | 2019-06-20 | Chugai Seiyaku Kabushiki Kaisha | Bispecific antibody exhibiting increased alternative fviii-cofactor-function activity |
EP3491022A1 (en) | 2016-07-29 | 2019-06-05 | Institut National de la Sante et de la Recherche Medicale (INSERM) | Antibodies targeting tumor associated macrophages and uses thereof |
KR20230079499A (en) | 2016-08-05 | 2023-06-07 | 추가이 세이야쿠 가부시키가이샤 | Composition for prophylaxis or treatment of il-8 related diseases |
JP2019530434A (en) | 2016-08-05 | 2019-10-24 | ジェネンテック, インコーポレイテッド | Multivalent and multi-epitope antibodies with agonist activity and methods of use |
EP3497129A1 (en) | 2016-08-08 | 2019-06-19 | H. Hoffnabb-La Roche Ag | Therapeutic and diagnostic methods for cancer |
WO2018029182A1 (en) | 2016-08-08 | 2018-02-15 | Ablynx N.V. | Il-6r single variable domain antibodies for treatment of il-6r related diseases |
WO2018031662A1 (en) | 2016-08-11 | 2018-02-15 | Genentech, Inc. | Pyrrolobenzodiazepine prodrugs and antibody conjugates thereof |
WO2018041989A1 (en) | 2016-09-02 | 2018-03-08 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for diagnosing and treating refractory celiac disease type 2 |
EP3510407A1 (en) | 2016-09-08 | 2019-07-17 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for diagnosing and treating nephrotic syndrome |
EP3512880A1 (en) | 2016-09-15 | 2019-07-24 | Ablynx NV | Immunoglobulin single variable domains directed against macrophage migration inhibitory factor |
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 |
JOP20190009A1 (en) | 2016-09-21 | 2019-01-27 | Alx Oncology Inc | Antibodies against signal-regulatory protein alpha and methods of use |
JP7051826B2 (en) | 2016-09-23 | 2022-04-11 | シーエスエル、リミテッド | Coagulation factor binding protein and its use |
EP3528838B1 (en) | 2016-09-23 | 2023-07-19 | F. Hoffmann-La Roche AG | Uses of il-13 antagonists for treating atopic dermatitis |
CN109843926B (en) | 2016-09-30 | 2023-01-20 | 豪夫迈·罗氏有限公司 | Bispecific antibodies against CD3 |
MA46366A (en) | 2016-09-30 | 2019-08-07 | Janssen Biotech Inc | SAFE AND EFFECTIVE PROCESS FOR TREATING PSORIASIS WITH A SPECIFIC ANTIBODY AGAINST IL-23 |
EP3522933B1 (en) | 2016-10-05 | 2021-12-15 | F. Hoffmann-La Roche AG | Methods for preparing antibody drug conjugates |
EP3522913A4 (en) | 2016-10-05 | 2020-10-28 | Acceleron Pharma Inc. | Alk4:actriib heteromultimers and uses thereof |
JP2019529509A (en) | 2016-10-05 | 2019-10-17 | アクセレロン ファーマ インコーポレーテッド | Compositions and methods for treating kidney disease |
CN110418851A (en) | 2016-10-06 | 2019-11-05 | 基因泰克公司 | The treatment of cancer and diagnostic method |
WO2018068201A1 (en) | 2016-10-11 | 2018-04-19 | Nanjing Legend Biotech Co., Ltd. | Single-domain antibodies and variants thereof against ctla-4 |
WO2018075564A1 (en) | 2016-10-17 | 2018-04-26 | University Of Maryland, College Park | Multispecific antibodies targeting human immunodeficiency virus and methods of using the same |
CN110366558A (en) | 2016-10-28 | 2019-10-22 | 班扬生物标记公司 | For the antibody and correlation technique of ubiquitin c-terminal hydrolase-l 1 (UCH-L1) and glial fibrillary acid protein (GFAP) |
US11555076B2 (en) | 2016-10-29 | 2023-01-17 | Genentech, Inc. | Anti-MIC antibodies and methods of use |
EP3538140A1 (en) | 2016-11-14 | 2019-09-18 | Institut National de la Sante et de la Recherche Medicale (INSERM) | Methods and pharmaceutical compositions for modulating stem cells proliferation or differentiation |
US11466094B2 (en) | 2016-11-15 | 2022-10-11 | Genentech, Inc. | Dosing for treatment with anti-CD20/anti-CD3 bispecific antibodies |
CN110177809B (en) | 2016-11-16 | 2023-11-03 | 埃博灵克斯股份有限公司 | T cell recruiting polypeptides capable of binding CD123 and TCR alpha/beta |
EP3541422A4 (en) | 2016-11-16 | 2020-05-06 | Janssen Biotech, Inc. | Method of treating psoriasis with anti-il-23 specific antibody |
WO2018091621A1 (en) | 2016-11-17 | 2018-05-24 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for increasing endogenous protein level |
TW201829463A (en) | 2016-11-18 | 2018-08-16 | 瑞士商赫孚孟拉羅股份公司 | Anti-hla-g antibodies and use thereof |
CN108367075B (en) | 2016-11-23 | 2022-08-09 | 免疫方舟医药技术股份有限公司 | 4-1BB binding proteins and uses thereof |
US11168139B2 (en) | 2016-11-28 | 2021-11-09 | Chugai Seiyaku Kabushiki Kaisha | Antigen-binding domain, and polypeptide including conveying section |
WO2018099968A1 (en) | 2016-11-29 | 2018-06-07 | Ablynx N.V. | Treatment of infection by respiratory syncytial virus (rsv) |
WO2018100190A1 (en) | 2016-12-02 | 2018-06-07 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for diagnosing renal cell carcinoma |
CN117820467A (en) | 2016-12-07 | 2024-04-05 | 基因泰克公司 | anti-TAU antibodies and methods of use |
PE20191135A1 (en) | 2016-12-07 | 2019-09-02 | Genentech Inc | ANTITAU ANTIBODIES AND METHODS OF USE |
EP3555620A1 (en) | 2016-12-13 | 2019-10-23 | H. Hoffnabb-La Roche Ag | Method to determine the presence of a target antigen in a tumor sample |
MX2019006866A (en) | 2016-12-16 | 2019-08-22 | Merck Patent Gmbh | Methods for the use of galectin 3 binding protein detected in the urine for monitoring the severity and progression of lupus nephritis. |
WO2018114754A1 (en) | 2016-12-19 | 2018-06-28 | F. Hoffmann-La Roche Ag | Combination therapy with targeted 4-1bb (cd137) agonists |
BR112019007267A2 (en) | 2016-12-20 | 2019-07-09 | Hoffmann La Roche | anti-cd20 / anti-cd3 bispecific antibody, pharmaceutical product, pharmaceutical composition comprising a anti-cd20 / anti-cd3 bispecific antibody, use of anti-cd20 / anti-cd3 bispecific antibody combination and a 4-1bb agonist and method of treatment or retardation of cancer progression in patients |
GB201621806D0 (en) | 2016-12-21 | 2017-02-01 | Philogen Spa | Immunocytokines with progressive activation mechanism |
EP3360898A1 (en) | 2017-02-14 | 2018-08-15 | Boehringer Ingelheim International GmbH | Bispecific anti-tnf-related apoptosis-inducing ligand receptor 2 and anti-cadherin 17 binding molecules for the treatment of cancer |
CN108261391B (en) | 2016-12-30 | 2022-03-01 | 江苏太平洋美诺克生物药业有限公司 | Stable pharmaceutical formulation comprising CD147 monoclonal antibody |
CN108261544B (en) | 2016-12-30 | 2023-05-05 | 江苏太平洋美诺克生物药业股份有限公司 | Stable pharmaceutical formulation comprising CD147 monoclonal antibody |
WO2018127473A1 (en) | 2017-01-03 | 2018-07-12 | F. Hoffmann-La Roche Ag | Bispecific antigen binding molecules comprising anti-4-1bb clone 20h4.9 |
WO2018134389A1 (en) | 2017-01-23 | 2018-07-26 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating infections |
AU2017396503A1 (en) | 2017-01-30 | 2019-07-11 | Janssen Biotech, Inc. | Anti-TNF antibodies, compositions, and methods for the treatment of active Psoriatic arthritis |
CN110234355B (en) | 2017-02-01 | 2021-11-09 | 浙江时迈药业有限公司 | Monomeric human IgG1Fc and bispecific antibodies |
AU2017398101A1 (en) | 2017-02-07 | 2019-08-01 | Janssen Biotech, Inc. | Anti-TNF antibodies, compositions, and methods for the treatment of active Ankylosing Spondylitis |
US11266745B2 (en) | 2017-02-08 | 2022-03-08 | Imaginab, Inc. | Extension sequences for diabodies |
JP7341060B2 (en) | 2017-02-10 | 2023-09-08 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Methods and pharmaceutical compositions for the treatment of cancer associated with MAPK pathway activation |
PE20191548A1 (en) | 2017-02-10 | 2019-10-24 | Genentech Inc | ANTIBODIES AGAINST TRYPTASE, COMPOSITIONS OF THESE AND USES OF THEM |
CN110573604A (en) | 2017-02-28 | 2019-12-13 | 非营利性组织佛兰芒综合大学生物技术研究所 | Means and methods for oral protein delivery |
MX2019010295A (en) | 2017-03-01 | 2019-11-21 | Genentech Inc | Diagnostic and therapeutic methods for cancer. |
WO2018158398A1 (en) | 2017-03-02 | 2018-09-07 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Antibodies having specificity to nectin-4 and uses thereof |
SG11201908547VA (en) | 2017-03-22 | 2019-10-30 | Genentech Inc | Hydrogel cross-linked hyaluronic acid prodrug compositions and methods |
AR111249A1 (en) | 2017-03-22 | 2019-06-19 | Genentech Inc | OPTIMIZED ANTIBODY COMPOSITIONS FOR THE TREATMENT OF OCULAR DISORDERS |
EP3600427A1 (en) | 2017-03-24 | 2020-02-05 | INSERM - Institut National de la Santé et de la Recherche Médicale | Methods and compositions for treating melanoma |
WO2018178030A1 (en) | 2017-03-27 | 2018-10-04 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating degenerative muscular and/or neurological conditions or diseases |
WO2018178029A1 (en) | 2017-03-27 | 2018-10-04 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating degenerative muscular and/or neurological conditions or diseases |
KR20190133723A (en) | 2017-03-27 | 2019-12-03 | 에프. 호프만-라 로슈 아게 | Improved antigen binding receptors |
RU2019133199A (en) | 2017-03-27 | 2021-04-28 | Ф. Хоффманн-Ля Рош Аг | IMPROVED ANTIGEN BINDING RECEPTOR FORMATS |
WO2018178078A1 (en) | 2017-03-28 | 2018-10-04 | INSERM (Institut National de la Santé et de la Recherche Médicale) | New tau species |
JP2020515543A (en) | 2017-03-28 | 2020-05-28 | ジェネンテック, インコーポレイテッド | How to treat neurodegenerative diseases |
WO2018178074A1 (en) | 2017-03-29 | 2018-10-04 | F. Hoffmann-La Roche Ag | Trimeric antigen binding molecules specific for a costimulatory tnf receptor |
JP7205995B2 (en) | 2017-03-29 | 2023-01-17 | 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 for co-stimulatory TNF receptors |
WO2018178055A1 (en) | 2017-03-29 | 2018-10-04 | F. Hoffmann-La Roche Ag | Bispecific antigen binding molecule for a costimulatory tnf receptor |
JP2020512348A (en) | 2017-03-30 | 2020-04-23 | デューク ユニバーシティ | Radiolabeled biomolecules and uses thereof |
CN110382525B (en) | 2017-04-03 | 2023-10-20 | 豪夫迈·罗氏有限公司 | Immunoconjugates |
PE20200010A1 (en) | 2017-04-03 | 2020-01-06 | Hoffmann La Roche | ANTIBODIES THAT JOIN STEAP-1 |
PE20191494A1 (en) | 2017-04-03 | 2019-10-21 | Hoffmann La Roche | IMMUNOCONJUGATES OF AN ANTI-PD-1 ANTIBODY WITH A MUTANT IL-2 OR IL-15 |
TWI704158B (en) | 2017-04-04 | 2020-09-11 | 瑞士商赫孚孟拉羅股份公司 | Novel bispecific antigen binding molecules capable of specific binding to cd40 and to fap |
CN116375876A (en) | 2017-04-05 | 2023-07-04 | 豪夫迈·罗氏有限公司 | Bispecific antibodies that specifically bind PD1 and LAG3 |
MA49034B1 (en) | 2017-04-05 | 2022-09-30 | Hoffmann La Roche | Anti-lag3 antibody |
CA3058290A1 (en) | 2017-04-18 | 2018-10-25 | Universite Libre De Bruxelles | Biomarkers and targets for proliferative diseases |
CN108728444A (en) | 2017-04-18 | 2018-11-02 | 长春华普生物技术股份有限公司 | Immunoregulation polynucleotide and its application |
WO2018195302A1 (en) | 2017-04-19 | 2018-10-25 | Bluefin Biomedicine, Inc. | Anti-vtcn1 antibodies and antibody drug conjugates |
MA49131A (en) | 2017-04-21 | 2020-03-25 | Hoffmann La Roche | USE OF KLK5 ANTAGONISTS FOR THE TREATMENT OF DISEASE |
CA3059468A1 (en) | 2017-04-27 | 2018-11-01 | Tesaro, Inc. | Antibody agents directed against lymphocyte activation gene-3 (lag-3) and uses thereof |
JP2020518638A (en) | 2017-05-05 | 2020-06-25 | アラコス インコーポレイテッド | Methods and compositions for treating allergic eye diseases |
JP2020519261A (en) | 2017-05-11 | 2020-07-02 | ブイアイビー ブイゼットダブリュVib Vzw | Glycosylation of variable immunoglobulin domains |
EP3625251A1 (en) | 2017-05-15 | 2020-03-25 | University Of Rochester | Broadly neutralizing anti-influenza monoclonal antibody and uses thereof |
EP3403649A1 (en) | 2017-05-16 | 2018-11-21 | Bayer Pharma Aktiengesellschaft | Inhibitors and antagonists of gpr84 for the treatment of endometriosis |
US20200171022A1 (en) | 2017-05-17 | 2020-06-04 | Inserm (Institut National De La Santé Et Da La Recherche Médicale) | Flt3 inhibitors for improving pain treatments by opioids |
EP3406253A1 (en) | 2017-05-24 | 2018-11-28 | Bayer Aktiengesellschaft | Inhibitors and antagonists of human pycr1 |
US10781265B2 (en) * | 2017-05-24 | 2020-09-22 | Development Center For Biotechnology | Humanized antibodies against Globo H and uses thereof in cancer treatments |
EP3409322A1 (en) | 2017-06-01 | 2018-12-05 | F. Hoffmann-La Roche AG | Treatment method |
BR112019024333A2 (en) | 2017-06-02 | 2020-07-28 | Merck Patent Gmbh | adamts binding immunoglobulins |
MX2019014397A (en) | 2017-06-02 | 2020-02-10 | Merck Patent Gmbh | Polypeptides binding adamts5, mmp13 and aggrecan. |
WO2018220225A1 (en) | 2017-06-02 | 2018-12-06 | Ablynx Nv | Aggrecan binding immunoglobulins |
AU2018278275A1 (en) | 2017-06-02 | 2019-12-19 | Ablynx N.V. | MMP13 binding immunoglobulins |
EP3634582A1 (en) | 2017-06-08 | 2020-04-15 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating hyperpigmentation disorders |
US20200096507A1 (en) | 2017-06-08 | 2020-03-26 | Inserm (Institut National De La Sante Et De La Recherche Medicale) | Chimeric receptor for use in whole-cell sensors for detecting analytes of interest |
US10947316B2 (en) * | 2017-06-15 | 2021-03-16 | Development Center For Biotechnology | Antibody-drug conjugates containing anti-Globo H antibodies and uses thereof |
US20210403573A1 (en) | 2017-06-22 | 2021-12-30 | INSERM (Institut National de la Santé et de la Recherche Médicale | Methods and pharmaceutical compositions for the treatment of fibrosis with agents capable of inhibiting the activation of mucosal-associated invariant t (mait) cells |
WO2019000223A1 (en) | 2017-06-27 | 2019-01-03 | Nanjing Legend Biotech Co., Ltd. | Chimeric antibody immune effctor cell engagers and methods of use thereof |
WO2019002548A1 (en) | 2017-06-29 | 2019-01-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Treating migraine by agonising trek1, trek2 or heteromers including them |
SG10201913147WA (en) | 2017-07-11 | 2020-02-27 | Compass Therapeutics Llc | Agonist antibodies that bind human cd137 and uses thereof |
KR102625929B1 (en) | 2017-07-19 | 2024-01-16 | 브이아이비 브이지더블유 | Serum albumin binder |
WO2019016310A1 (en) | 2017-07-20 | 2019-01-24 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating cancers |
CN111492245A (en) | 2017-07-21 | 2020-08-04 | 基因泰克公司 | Methods of treatment and diagnosis of cancer |
WO2019020480A1 (en) | 2017-07-24 | 2019-01-31 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Antibodies and peptides to treat hcmv related diseases |
WO2019020593A1 (en) | 2017-07-25 | 2019-01-31 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for modulating monocytopoiesis |
AU2018308930A1 (en) | 2017-07-28 | 2020-02-06 | F. Hoffmann-La Roche Ag | Bispecific antibody formulation |
EP3589658A1 (en) | 2017-08-03 | 2020-01-08 | Alector LLC | Anti-cd33 antibodies and methods of use thereof |
EP3444275A1 (en) | 2017-08-16 | 2019-02-20 | Exiris S.r.l. | Monoclonal antibody anti-fgfr4 |
WO2019036855A1 (en) | 2017-08-21 | 2019-02-28 | Adagene Inc. | Anti-cd137 molecules and use thereof |
US20200197547A1 (en) | 2017-08-30 | 2020-06-25 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Anti-mesothelin radiolabelled single domain antibodies suitable for the imaging and treatment of cancers |
EP3457139A1 (en) | 2017-09-19 | 2019-03-20 | Promise Advanced Proteomics | Antibody-like peptides for quantifying therapeutic antibodies |
WO2019057742A1 (en) | 2017-09-20 | 2019-03-28 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for modulating autophagy |
JP7382922B2 (en) | 2017-09-20 | 2023-11-17 | 中外製薬株式会社 | Dosing regimen for combination therapy using PD-1 system binding antagonists and GPC3 targeting agents |
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 |
WO2019077092A1 (en) | 2017-10-20 | 2019-04-25 | F. Hoffmann-La Roche Ag | Method for generating multispecific antibodies from monospecific antibodies |
WO2019081456A1 (en) | 2017-10-24 | 2019-05-02 | Bayer Aktiengesellschaft | Use of activators and stimulators of sgc comprising a beta2 subunit |
AU2018358883A1 (en) | 2017-10-30 | 2020-04-23 | F. Hoffmann-La Roche Ag | Method for in vivo generation of multispecific antibodies from monospecific antibodies |
EP3704160A1 (en) | 2017-10-31 | 2020-09-09 | VIB vzw | Novel antigen-binding chimeric proteins and methods and uses thereof |
US11718679B2 (en) | 2017-10-31 | 2023-08-08 | Compass Therapeutics Llc | CD137 antibodies and PD-1 antagonists and uses thereof |
CN111315781A (en) | 2017-11-01 | 2020-06-19 | 豪夫迈·罗氏有限公司 | Combination therapy with a targeted OX40 agonist |
EP3703746A1 (en) | 2017-11-01 | 2020-09-09 | F. Hoffmann-La Roche AG | Novel tnf family ligand trimer-containing antigen binding molecules |
MA50505A (en) | 2017-11-01 | 2020-09-09 | Hoffmann La Roche | 2 + 1 BISPECIFIC ANTIBODIES (CONTORSBODIES) |
JP2021500930A (en) | 2017-11-01 | 2021-01-14 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | COMP Body-Multivalent Target Binding Substance |
CN111213059B (en) | 2017-11-06 | 2024-01-09 | 豪夫迈·罗氏有限公司 | Diagnostic and therapeutic methods for cancer |
WO2019094595A2 (en) | 2017-11-09 | 2019-05-16 | Pinteon Therapeutics Inc. | Methods and compositions for the generation and use of humanized conformation-specific phosphorylated tau antibodies |
EP3710486A1 (en) | 2017-11-15 | 2020-09-23 | Novo Nordisk A/S | Factor x binders enhancing fx activation |
WO2019100052A2 (en) | 2017-11-20 | 2019-05-23 | Compass Therapeutics Llc | Cd137 antibodies and tumor antigen-targeting antibodies and uses thereof |
WO2019101995A1 (en) | 2017-11-27 | 2019-05-31 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for cardiac regeneration |
AU2018377783A1 (en) | 2017-11-28 | 2020-06-11 | Chugai Seiyaku Kabushiki Kaisha | Polypeptide including antigen-binding domain and carrying section |
PL3717011T3 (en) | 2017-11-29 | 2023-03-27 | Csl Limited | Method of treating or preventing ischemia-reperfusion injury |
MA51208A (en) | 2017-12-01 | 2020-10-07 | Seattle Genetics Inc | CD47 ANTIBODIES (MASKED) AND THEIR USES FOR CANCER TREATMENT |
MX2020005640A (en) | 2017-12-01 | 2020-08-20 | Seattle Genetics Inc | Humanized anti-liv1 antibodies for the treatment of breast cancer. |
MX2020006125A (en) | 2017-12-14 | 2020-08-24 | Hoffmann La Roche | Use of a cea cd3 bispecific antibody and a pd-1 axis binding antagonist in a dosage regime to treat cancer. |
MX2020006372A (en) | 2017-12-19 | 2020-09-03 | Univ Rockefeller | HUMAN IgG Fc DOMAIN VARIANTS WITH IMPROVED EFFECTOR FUNCTION. |
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 |
PE20201149A1 (en) | 2017-12-21 | 2020-10-26 | Hoffmann La Roche | HLA-A2 / WT1 BINDING ANTIBODIES |
JP2021508246A (en) | 2017-12-21 | 2021-03-04 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | CAR-T cell assay for specificity testing of novel antigen binding moiety |
JP7394058B2 (en) | 2017-12-21 | 2023-12-07 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Universal reporter cell assay for specificity testing of novel antigen-binding moieties |
US20190211098A1 (en) | 2017-12-22 | 2019-07-11 | Genentech, Inc. | Use of pilra binding agents for treatment of a disease |
KR20200104333A (en) | 2017-12-28 | 2020-09-03 | 난징 레전드 바이오테크 씨오., 엘티디. | Single-domain antibodies to TIGIT and variants thereof |
WO2019133512A1 (en) | 2017-12-29 | 2019-07-04 | Alector Llc | Anti-tmem106b antibodies and methods of use thereof |
WO2019134946A1 (en) | 2018-01-04 | 2019-07-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma resistant |
EP3740507A4 (en) | 2018-01-15 | 2022-08-24 | Nanjing Legend Biotech Co., Ltd. | Single-domain antibodies and variants thereof against pd-1 |
WO2019143636A1 (en) | 2018-01-16 | 2019-07-25 | Lakepharma, Inc. | Bispecific antibody that binds cd3 and another target |
US20210054064A1 (en) | 2018-01-24 | 2021-02-25 | INSERM (Institut National de la Santé et de la Recherche Médicale | Antagonists of il-33 for use in methods for preventing ischemia reperfusion injusry in an organ |
WO2019149715A1 (en) | 2018-01-31 | 2019-08-08 | F. Hoffmann-La Roche Ag | Stabilized immunoglobulin domains |
WO2019152715A1 (en) | 2018-01-31 | 2019-08-08 | Alector Llc | Anti-ms4a4a antibodies and methods of use thereof |
WO2019149716A1 (en) | 2018-01-31 | 2019-08-08 | F. Hoffmann-La Roche Ag | Bispecific antibodies comprising an antigen-binding site binding to lag3 |
WO2019148444A1 (en) | 2018-02-02 | 2019-08-08 | Adagene Inc. | Anti-ctla4 antibodies and methods of making and using the same |
WO2019148445A1 (en) | 2018-02-02 | 2019-08-08 | Adagene Inc. | Precision/context-dependent activatable antibodies, and methods of making and using the same |
WO2019150309A1 (en) | 2018-02-02 | 2019-08-08 | Hammack Scott | Modulators of gpr68 and uses thereof for treating and preventing diseases |
US20210221912A1 (en) | 2018-02-06 | 2021-07-22 | Ablynx N.V. | Methods of treating initial episode of ttp with immunoglobulin single variable domains |
MX2020008289A (en) | 2018-02-08 | 2020-09-25 | Genentech Inc | Bispecific antigen-binding molecules and methods of use. |
WO2019157358A1 (en) | 2018-02-09 | 2019-08-15 | Genentech, Inc. | Therapeutic and diagnostic methods for mast cell-mediated inflammatory diseases |
TWI829667B (en) | 2018-02-09 | 2024-01-21 | 瑞士商赫孚孟拉羅股份公司 | Antibodies binding to gprc5d |
JP7384811B2 (en) | 2018-02-16 | 2023-11-21 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Methods and compositions for treating vitiligo |
JP7391027B2 (en) | 2018-02-26 | 2023-12-04 | ジェネンテック, インコーポレイテッド | Medication for treatment with anti-TIGIT and anti-PD-L1 antagonist antibodies |
JP2021514648A (en) | 2018-03-01 | 2021-06-17 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | Specificity assay for novel target antigen binding moieties |
JP2021514649A (en) | 2018-03-01 | 2021-06-17 | ブレイエ・ユニバージテイト・ブリュッセルVrije Universiteit Brussel | Human PD-L1 binding immunoglobulin |
AU2019232624A1 (en) | 2018-03-05 | 2020-09-10 | Janssen Biotech, Inc. | Methods of treating Crohn's disease with anti-IL23 specific antibody |
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 |
BR112020015568A2 (en) | 2018-03-13 | 2020-12-29 | F. Hoffmann-La Roche Ag | 4-1BB AGONIST (CD137), PHARMACEUTICAL PRODUCT, PHARMACEUTICAL COMPOSITION, USE OF A COMBINATION OF A 4-1BB AGONIST AND METHOD TO TREAT OR DELAY CANCER PROGRESSION |
US20200040103A1 (en) | 2018-03-14 | 2020-02-06 | Genentech, Inc. | Anti-klk5 antibodies and methods of use |
RU2020128013A (en) | 2018-03-14 | 2022-04-15 | Бейцзин Сюаньи Фармасайенсиз Ко., Лтд. | ANTIBODIES AGAINST CLAUDIN 18.2 |
KR20200132938A (en) | 2018-03-15 | 2020-11-25 | 추가이 세이야쿠 가부시키가이샤 | Anti-dengue virus antibodies with cross-reactivity against Zika virus and methods of use |
EP3768720A4 (en) | 2018-03-20 | 2022-01-05 | Wuxi Biologics Ireland Limited | Novel anti-lag-3 antibody polypeptide |
MA52091A (en) | 2018-03-21 | 2021-01-27 | Alx Oncology Inc | ALPHA SIGNAL REGULATING PROTEIN ANTIBODIES AND METHODS OF USE |
PL3768701T3 (en) | 2018-03-23 | 2024-02-19 | Université Libre de Bruxelles | Wnt signaling agonist molecules |
SG11202009542PA (en) | 2018-03-29 | 2020-10-29 | Genentech Inc | Modulating lactogenic activity in mammalian cells |
CA3093034A1 (en) | 2018-03-30 | 2019-10-03 | Nanjing Legend Biotech Co., Ltd. | Single-domain antibodies against lag-3 and uses thereof |
JP7104458B2 (en) | 2018-04-02 | 2022-07-21 | 上海博威生物医薬有限公司 | Lymphocyte activation gene-3 (LAG-3) -binding antibody and its use |
TW202011029A (en) | 2018-04-04 | 2020-03-16 | 美商建南德克公司 | Methods for detecting and quantifying FGF21 |
EP3775902B1 (en) | 2018-04-04 | 2023-02-22 | F. Hoffmann-La Roche AG | Diagnostic assays to detect tumor antigens in cancer patients |
CN112424601A (en) | 2018-04-04 | 2021-02-26 | 豪夫迈·罗氏有限公司 | Diagnostic assay for detecting tumor antigens in cancer patients |
WO2019193375A1 (en) | 2018-04-04 | 2019-10-10 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of fzd7 inhibitors for the treatment of retinal neovascularization |
EP3552631A1 (en) | 2018-04-10 | 2019-10-16 | Inatherys | Antibody-drug conjugates and their uses for the treatment of cancer |
CR20200459A (en) | 2018-04-13 | 2020-11-11 | Hoffmann La Roche | Her2-targeting antigen binding molecules comprising 4-1bbl |
AR115052A1 (en) | 2018-04-18 | 2020-11-25 | Hoffmann La Roche | MULTI-SPECIFIC ANTIBODIES AND THE USE OF THEM |
AR114789A1 (en) | 2018-04-18 | 2020-10-14 | Hoffmann La Roche | ANTI-HLA-G ANTIBODIES AND THE USE OF THEM |
WO2019207030A1 (en) | 2018-04-26 | 2019-10-31 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for predicting a response with an immune checkpoint inhibitor in a patient suffering from a lung cancer |
WO2019213384A1 (en) | 2018-05-03 | 2019-11-07 | University Of Rochester | Anti-influenza neuraminidase monoclonal antibodies and uses thereof |
FI3794024T3 (en) | 2018-05-14 | 2023-08-10 | Werewolf Therapeutics Inc | Activatable interleukin-2 polypeptides and methods of use thereof |
JP2021524756A (en) | 2018-05-14 | 2021-09-16 | ウェアウルフ セラピューティクス, インコーポレイテッド | Activateable cytokine polypeptides and how to use them |
AU2019269066B2 (en) | 2018-05-18 | 2022-10-06 | F. Hoffmann-La Roche Ag | Targeted intracellular delivery of large nucleic acids |
EP3569618A1 (en) | 2018-05-19 | 2019-11-20 | Boehringer Ingelheim International GmbH | Antagonizing cd73 antibody |
US11319373B2 (en) | 2018-05-25 | 2022-05-03 | Alector Llc | Anti-SIRPA antibodies and methods of use thereof |
EP3816182A4 (en) | 2018-05-30 | 2022-07-13 | Chugai Seiyaku Kabushiki Kaisha | Ligand-binding molecule containing single domain antibody |
MX2020012997A (en) | 2018-06-01 | 2021-03-29 | Eisai R&D Man Co Ltd | Splicing modulator antibody-drug conjugates and methods of use. |
EP3801523A2 (en) | 2018-06-01 | 2021-04-14 | Eisai R&D Management Co., Ltd. | Methods of using splicing modulators |
WO2019235426A1 (en) | 2018-06-04 | 2019-12-12 | 中外製薬株式会社 | Antigen-binding molecule showing changed half-life in cytoplasm |
EP3801613A1 (en) | 2018-06-04 | 2021-04-14 | Bayer Aktiengesellschaft | Inhibitors of shp2 |
TW202016151A (en) | 2018-06-09 | 2020-05-01 | 德商百靈佳殷格翰國際股份有限公司 | Multi-specific binding proteins for cancer treatment |
KR20210035805A (en) | 2018-06-15 | 2021-04-01 | 플래그쉽 파이어니어링 이노베이션스 브이, 인크. | Increased immune activity through regulation of postcellular signaling factors |
US20210277118A1 (en) | 2018-06-21 | 2021-09-09 | Daiichi Sankyo Company, Limited | Compositions including cd3 antigen binding fragments and uses thereof |
TWI819011B (en) | 2018-06-23 | 2023-10-21 | 美商建南德克公司 | Methods of treating lung cancer with a pd-1 axis binding antagonist, a platinum agent, and a topoisomerase ii inhibitor |
BR112020026819A2 (en) | 2018-06-29 | 2021-04-20 | Alector Llc | isolated antibodies, nucleic acid, vector, host cells, method of producing an antibody, pharmaceutical composition, methods for treating cancer and for treating a disease and uses of an antibody |
CN112955465A (en) | 2018-07-03 | 2021-06-11 | 马伦戈治疗公司 | anti-TCR antibody molecules and uses thereof |
JP2021528988A (en) | 2018-07-04 | 2021-10-28 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | New bispecific agonist 4-1BB antigen-binding molecule |
WO2020008083A1 (en) | 2018-07-05 | 2020-01-09 | Consejo Superior De Investigaciones Científicas | Therapeutic target in chemokine receptors for the screening of compounds useful for the treatment of pathological processes involving chemokine signaling |
TWI809147B (en) | 2018-07-13 | 2023-07-21 | 美商阿列克特有限責任公司 | Anti-sortilin antibodies and methods of use thereof |
WO2020016838A2 (en) | 2018-07-18 | 2020-01-23 | Janssen Biotech, Inc. | Sustained response predictors after treatment with anti-il23 specific antibody |
JP2021530502A (en) | 2018-07-18 | 2021-11-11 | ジェネンテック, インコーポレイテッド | How to Treat Lung Cancer with PD-1 Axial Binding Antagonists, Antimetabolites, and Platinums |
CA3106114A1 (en) | 2018-07-20 | 2020-01-23 | Surface Oncology, Inc. | Anti-cd112r compositions and methods |
CN113286819A (en) | 2018-08-01 | 2021-08-20 | 感应检查疗法公司 | anti-BTN 3A antibodies and their use in treating cancer or infectious disorders |
EP3831854A4 (en) | 2018-08-03 | 2022-05-04 | Chugai Seiyaku Kabushiki Kaisha | Antigen-binding molecule containing two antigen-binding domains that are linked to each other |
JP2021533149A (en) | 2018-08-08 | 2021-12-02 | ジェネンテック, インコーポレイテッド | Use of tryptophan derivatives and L-methionine for protein formulations |
WO2020032230A1 (en) | 2018-08-10 | 2020-02-13 | 中外製薬株式会社 | Anti-cd137 antigen-binding molecule and utilization thereof |
TW202021618A (en) | 2018-08-17 | 2020-06-16 | 美商23與我有限公司 | Anti-il1rap antibodies and methods of use thereof |
WO2020041360A1 (en) | 2018-08-21 | 2020-02-27 | Quidel Corporation | Dbpa antibodies and uses thereof |
JP2021534797A (en) | 2018-08-31 | 2021-12-16 | アレクトル エルエルシー | Anti-CD33 antibody and its usage |
GB201814281D0 (en) | 2018-09-03 | 2018-10-17 | Femtogenix Ltd | Cytotoxic agents |
CN112654397A (en) | 2018-09-05 | 2021-04-13 | 国家医疗保健研究所 | Methods and compositions for treating asthma and allergic diseases |
GB2576914A (en) | 2018-09-06 | 2020-03-11 | Kymab Ltd | Antigen-binding molecules comprising unpaired variable domains produced in mammals |
EP3849545A1 (en) | 2018-09-10 | 2021-07-21 | Institut National de la Santé et de la Recherche Médicale (INSERM) | Methods for the treatment of neurofibromatosis |
EP3849602A1 (en) | 2018-09-10 | 2021-07-21 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Combination of her2/neu antibody with heme for treating cancer |
EP3850008A1 (en) | 2018-09-10 | 2021-07-21 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of an inhibitor of ntsr1 activation or expression for preventing weight loss, muscle loss, and protein blood level decrease in subjects in need thereof |
WO2020058201A1 (en) | 2018-09-17 | 2020-03-26 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of inhibitors of phosphatase activity of soluble epoxide for the treatment of cardiometabolic diseases |
TW202023542A (en) | 2018-09-18 | 2020-07-01 | 瑞士商赫孚孟拉羅股份公司 | Use of a cathepsin s inhibitor against the formation of anti-drug antibodies |
AU2019342099A1 (en) | 2018-09-19 | 2021-04-08 | Genentech, Inc. | Therapeutic and diagnostic methods for bladder cancer |
WO2020058372A1 (en) | 2018-09-19 | 2020-03-26 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical composition for the treatment of cancers resistant to immune checkpoint therapy |
EP3857230B1 (en) | 2018-09-21 | 2023-06-07 | F. Hoffmann-La Roche AG | Diagnostic methods for triple-negative breast cancer |
EP3883606B9 (en) | 2018-09-24 | 2023-10-04 | Janssen Biotech, Inc. | Safe and effective method of treating ulcerative colitis with anti-il12/il23 antibody |
WO2020064702A1 (en) | 2018-09-25 | 2020-04-02 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of antagonists of th17 cytokines for the treatment of bronchial remodeling in patients suffering from allergic asthma |
KR20210087027A (en) | 2018-09-27 | 2021-07-09 | 실리오 디벨럽먼트, 인크. | Masked cytokine polypeptide |
WO2020070062A1 (en) | 2018-10-01 | 2020-04-09 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of tim-3 inhibitors for the treatment of exacerbations in patients suffering from severe asthma |
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 |
KR20210069675A (en) | 2018-10-01 | 2021-06-11 | 에프. 호프만-라 로슈 아게 | Bispecific antigen binding molecule comprising anti-FAP clone 212 |
JP2022512626A (en) | 2018-10-04 | 2022-02-07 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Methods and pharmaceutical compositions for the treatment of mucosal inflammatory diseases |
EP3860653A1 (en) | 2018-10-05 | 2021-08-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and systems for controlling the agonistic properties of antibody variable domains by light |
EP3636657A1 (en) | 2018-10-08 | 2020-04-15 | Ablynx N.V. | Chromatography-free antibody purification method |
JP2022512648A (en) | 2018-10-09 | 2022-02-07 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Use of αV-integrin (CD51) inhibitors for the treatment of myocardial fibrosis |
TW202028244A (en) | 2018-10-09 | 2020-08-01 | 美商建南德克公司 | Methods and systems for determining synapse formation |
WO2020081493A1 (en) | 2018-10-16 | 2020-04-23 | Molecular Templates, Inc. | Pd-l1 binding proteins |
WO2020081767A1 (en) | 2018-10-18 | 2020-04-23 | Genentech, Inc. | Diagnostic and therapeutic methods for sarcomatoid kidney cancer |
WO2020079162A1 (en) | 2018-10-18 | 2020-04-23 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for inducing full ablation of hematopoiesis |
AU2019365238A1 (en) | 2018-10-24 | 2021-05-13 | F. Hoffmann-La Roche Ag | Conjugated chemical inducers of degradation and methods of use |
TW202031899A (en) | 2018-11-05 | 2020-09-01 | 美商建南德克公司 | Methods of producing two chain proteins in prokaryotic host cells |
MX2021005594A (en) | 2018-11-13 | 2021-10-22 | Compass Therapeutics Llc | Multispecific binding constructs against checkpoint molecules and uses thereof. |
GB201818477D0 (en) | 2018-11-13 | 2018-12-26 | Emstopa Ltd | Tissue plasminogen activator antibodies and method of use thereof |
JP7463366B2 (en) | 2018-11-20 | 2024-04-08 | タケダ ワクチン,インコーポレイテッド | Novel anti-Zika virus antibodies and uses thereof |
WO2020104479A1 (en) | 2018-11-20 | 2020-05-28 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating cancers and resistant cancers with anti transferrin receptor 1 antibodies |
CN113395979A (en) | 2018-11-20 | 2021-09-14 | 詹森生物科技公司 | Safe and effective methods for treating psoriasis with anti-IL-23 specific antibodies |
MX2021006573A (en) | 2018-12-06 | 2021-07-15 | Genentech Inc | Combination therapy of diffuse large b-cell lymphoma comprising an anti-cd79b immunoconjugates, an alkylating agent and an anti-cd20 antibody. |
WO2020115261A1 (en) | 2018-12-07 | 2020-06-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
JP2022513198A (en) | 2018-12-10 | 2022-02-07 | ジェネンテック, インコーポレイテッド | Photocrosslinkable peptide for site-specific conjugation to Fc-containing proteins |
WO2020120592A1 (en) | 2018-12-12 | 2020-06-18 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for predicting and treating melanoma |
CA3114931A1 (en) | 2018-12-13 | 2020-06-18 | Eisai R&D Management Co., Ltd. | Herboxidiene antibody-drug conjugates and methods of use |
EP3894438A1 (en) | 2018-12-13 | 2021-10-20 | INSERM (Institut National de la Santé et de la Recherche Médicale) | New anti tau svqivykpv epitope single domain antibody |
EP3897722A4 (en) | 2018-12-18 | 2022-09-14 | Janssen Biotech, Inc. | Safe and effective method of treating lupus with anti-il12/il23 antibody |
AR117327A1 (en) | 2018-12-20 | 2021-07-28 | 23Andme Inc | ANTI-CD96 ANTIBODIES AND METHODS OF USE OF THEM |
EP3898984A1 (en) | 2018-12-21 | 2021-10-27 | Genentech, Inc. | Methods of producing polypeptides using a cell line resistant to apoptosis |
EP3898673A1 (en) | 2018-12-21 | 2021-10-27 | 23Andme, Inc. | Anti-il-36 antibodies and methods of use thereof |
WO2020127885A1 (en) | 2018-12-21 | 2020-06-25 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Compositions for treating cancers and resistant cancers |
TW202030204A (en) | 2018-12-21 | 2020-08-16 | 瑞士商赫孚孟拉羅股份公司 | Tumor-targeted superagonistic cd28 antigen binding molecules |
JP7061733B2 (en) | 2018-12-21 | 2022-04-28 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Antibodies that bind to CD3 |
AU2019410073A1 (en) | 2018-12-21 | 2021-06-10 | F. Hoffmann-La Roche Ag | Tumor-targeted agonistic CD28 antigen binding molecules |
CA3123050A1 (en) | 2018-12-26 | 2020-07-02 | City Of Hope | Activatable masked anti-ctla4 binding proteins |
JP2022515543A (en) | 2018-12-30 | 2022-02-18 | エフ.ホフマン-ラ ロシュ アーゲー | Anti-rabbit CD19 antibody and how to use |
CN113574386A (en) | 2019-01-03 | 2021-10-29 | 国家医疗保健研究所 | Methods and pharmaceutical compositions for enhancing CD8+ T cell dependent immune responses in cancer patients |
WO2020150152A1 (en) | 2019-01-14 | 2020-07-23 | Genentech, Inc. | Methods of treating cancer with a pd-1 axis binding antagonist and an rna vaccine |
WO2020148651A1 (en) | 2019-01-15 | 2020-07-23 | Janssen Biotech, Inc. | Anti-tnf antibody compositions and methods for the treatment of juvenile idiopathic arthritis |
MA54814A (en) | 2019-01-23 | 2021-12-01 | Janssen Biotech Inc | ANTI-TNF ANTIBODY COMPOSITIONS FOR USE IN METHODS OF TREATING PSORIATIC ARTHRITIS |
SG11202106713UA (en) | 2019-01-23 | 2021-07-29 | Genentech Inc | Methods of producing multimeric proteins in eukaryotic host cells |
WO2020153467A1 (en) | 2019-01-24 | 2020-07-30 | 中外製薬株式会社 | Novel cancer antigens and antibodies of said antigens |
EP3917957A1 (en) | 2019-01-28 | 2021-12-08 | Maple Biotech LLC | Psmp antagonists for use in treatment of fibrotic disease of the lung, kidney or liver |
GB201901197D0 (en) | 2019-01-29 | 2019-03-20 | Femtogenix Ltd | G-A Crosslinking cytotoxic agents |
EP3921031A1 (en) | 2019-02-04 | 2021-12-15 | Institut National de la Santé et de la Recherche Médicale (INSERM) | Methods and compositions for modulating blood-brain barrier |
EP3921443A1 (en) | 2019-02-08 | 2021-12-15 | F. Hoffmann-La Roche AG | Diagnostic and therapeutic methods for cancer |
EP3927831A1 (en) | 2019-02-18 | 2021-12-29 | ATB Therapeutics | Method of producing a binder-toxin fusion protein in a plant cell or a whole plant |
WO2020169707A1 (en) | 2019-02-21 | 2020-08-27 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Foxo1 inhibitor for use in the treatment of latent virus infection |
WO2020169698A1 (en) | 2019-02-21 | 2020-08-27 | F. Hoffmann-La Roche Ag | Sensitization of cancer cells to tnf by bet inhibition |
CA3130695A1 (en) | 2019-02-27 | 2020-09-03 | Genentech, Inc. | Dosing for treatment with anti-tigit and anti-cd20 or anti-cd38 antibodies |
AU2020231308A1 (en) | 2019-03-01 | 2021-08-19 | Allogene Therapeutics, Inc. | DLL3 targeting chimeric antigen receptors and binding agents |
EP3935391B1 (en) | 2019-03-05 | 2024-04-24 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Biomarkers for renal cell carcinoma |
CN113474653A (en) | 2019-03-08 | 2021-10-01 | 豪夫迈·罗氏有限公司 | Method for detecting and quantifying membrane-associated proteins on extracellular vesicles |
WO2020183270A1 (en) | 2019-03-14 | 2020-09-17 | Janssen Biotech, Inc. | Methods for producing anti-tnf antibody compositions |
US20200291107A1 (en) | 2019-03-14 | 2020-09-17 | Janssen Biotech, Inc. | Manufacturing Methods for Producing Anti-IL12/IL23 Antibody Compositions |
MA55282A (en) | 2019-03-14 | 2022-01-19 | Janssen Biotech Inc | MANUFACTURING METHODS FOR THE PRODUCTION OF ANTI-TNF ANTIBODY COMPOSITIONS |
SG11202109424RA (en) | 2019-03-14 | 2021-09-29 | Genentech Inc | Treatment of cancer with her2xcd3 bispecific antibodies in combination with anti-her2 mab |
US20220153829A1 (en) | 2019-03-14 | 2022-05-19 | Janssen Biotech, Inc. | Methods for Producing Anti-TNF Antibody Compositions |
CN113631575A (en) | 2019-03-15 | 2021-11-09 | 笛卡尔疗法股份有限公司 | anti-BCMA chimeric antigen receptors |
CN113853385A (en) | 2019-03-18 | 2021-12-28 | 詹森生物科技公司 | Methods of treating psoriasis in pediatric subjects with anti-IL 12/IL23 antibodies |
EP3943108A4 (en) | 2019-03-19 | 2023-01-04 | Chugai Seiyaku Kabushiki Kaisha | Antigen-binding molecule containing antigen-binding domain of which binding activity to antigen is changed depending on mta, and library for obtaining said antigen-binding domain |
EP3947446A1 (en) | 2019-03-25 | 2022-02-09 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Treatment of taupathy disorders by targeting new tau species |
EP3946330A1 (en) | 2019-03-29 | 2022-02-09 | Institut National de la Santé et de la Recherche Médicale (INSERM) | Methods for the treatment of keloid, hypertrophic scars and/or hyperpigmentation disorders |
CA3130862A1 (en) | 2019-03-29 | 2020-10-08 | Genentech, Inc. | Modulators of cell surface protein interactions and methods and compositions related to same |
US20220177978A1 (en) | 2019-04-02 | 2022-06-09 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods of predicting and preventing cancer in patients having premalignant lesions |
US20220175815A1 (en) | 2019-04-03 | 2022-06-09 | Orega Biotech | Combination therapies based on pd1 and il-17b inhibitors |
WO2020201195A2 (en) | 2019-04-04 | 2020-10-08 | Bayer Aktiengesellschaft | Agonists of adiponectin |
SG11202109901TA (en) | 2019-04-09 | 2021-10-28 | Hospital For Special Surgery | Protein binders for irhom2 |
CN113677403A (en) | 2019-04-12 | 2021-11-19 | 豪夫迈·罗氏有限公司 | Bispecific antigen binding molecules comprising lipocalin muteins |
CN113692414A (en) | 2019-04-12 | 2021-11-23 | 豪夫迈·罗氏有限公司 | Cancer treatment using CEA CD3 bispecific antibodies and Wnt signaling inhibitors |
CN114364703A (en) | 2019-04-19 | 2022-04-15 | 豪夫迈·罗氏有限公司 | Anti-merk antibodies and methods of use thereof |
JP2022529741A (en) | 2019-04-26 | 2022-06-23 | アロジーン セラピューティクス,インコーポレイテッド | Method for Producing Allogeneic CAR T Cells |
WO2020221796A1 (en) | 2019-04-30 | 2020-11-05 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
EP3962947A2 (en) | 2019-05-03 | 2022-03-09 | F. Hoffmann-La Roche AG | Methods of treating cancer with an anti-pd-l1 antibody |
EP3962493A2 (en) | 2019-05-03 | 2022-03-09 | Flagship Pioneering Innovations V, Inc. | Methods of modulating immune activity/level of irf or sting or of treating cancer, comprising the administration of a sting modulator and/or purinergic receptor modulator or postcellular signaling factor |
MX2021013825A (en) | 2019-05-14 | 2022-01-18 | Genentech Inc | Methods of using anti-cd79b immunoconjugates to treat follicular lymphoma. |
CN114450022A (en) | 2019-05-14 | 2022-05-06 | 狼人治疗公司 | Separation fraction and method of use thereof |
JP2022534020A (en) | 2019-05-23 | 2022-07-27 | ヤンセン バイオテツク,インコーポレーテツド | Methods of treating inflammatory bowel disease with combination therapy of antibodies against IL-23 and TNF-alpha |
US20220228116A1 (en) | 2019-05-28 | 2022-07-21 | Vib Vzw | Cd8+ t-cells lacking plexins and their application in cancer treatment |
EP3976650A1 (en) | 2019-05-28 | 2022-04-06 | Vib Vzw | Cancer treatment by targeting plexins in the immune compartment |
WO2020245677A1 (en) | 2019-06-03 | 2020-12-10 | Janssen Biotech, Inc. | Anti-tnf antibodies, compositions, and methods for the treatment of active ankylosing spondylitis |
WO2020245676A1 (en) | 2019-06-03 | 2020-12-10 | Janssen Biotech, Inc. | Anti-tnf antibody compositions, and methods for the treatment of psoriatic arthritis |
JPWO2020246567A1 (en) | 2019-06-05 | 2020-12-10 | ||
KR20220017430A (en) | 2019-06-05 | 2022-02-11 | 추가이 세이야쿠 가부시키가이샤 | Antibody Cleavage Site Binding Molecules |
JPWO2020246617A1 (en) | 2019-06-07 | 2020-12-10 | ||
US20200392229A1 (en) | 2019-06-11 | 2020-12-17 | Alector Llc | Methods of use of anti-sortilin antibodies |
EP3986453A1 (en) | 2019-06-20 | 2022-04-27 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Anti- protease nexin-1 conformational single domain antibodies and uses thereof |
CN114127123A (en) | 2019-06-26 | 2022-03-01 | 豪夫迈·罗氏有限公司 | Fusion of CEA-binding antibodies to 4-1BBL |
CN114531878A (en) | 2019-06-27 | 2022-05-24 | 豪夫迈·罗氏有限公司 | Novel ICOS antibodies and tumor-targeted antigen-binding molecules comprising same |
EP3994169A1 (en) | 2019-07-02 | 2022-05-11 | F. Hoffmann-La Roche AG | Immunoconjugates comprising a mutant interleukin-2 and an anti-cd8 antibody |
AR119393A1 (en) | 2019-07-15 | 2021-12-15 | Hoffmann La Roche | ANTIBODIES THAT BIND NKG2D |
AU2020318975A1 (en) | 2019-07-22 | 2022-03-17 | Seagen Inc. | Humanized anti-LIV1 antibodies for the treatment of cancer |
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 |
US11667699B2 (en) | 2019-07-31 | 2023-06-06 | Alector Llc | Anti-MS4A4A antibodies and methods of use thereof |
CN114401743A (en) | 2019-08-02 | 2022-04-26 | 法国国家健康和医学研究院 | Use of neutralizing granzyme B for providing cardioprotection in a subject who has experienced a myocardial infarction |
WO2021028752A1 (en) | 2019-08-15 | 2021-02-18 | Janssen Biotech, Inc. | Anti-tfn antibodies for treating type i diabetes |
WO2021048292A1 (en) | 2019-09-11 | 2021-03-18 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
CN114340675A (en) | 2019-09-12 | 2022-04-12 | 豪夫迈·罗氏有限公司 | Compositions and methods for treating lupus nephritis |
US11918649B2 (en) | 2019-09-18 | 2024-03-05 | Molecular Templates, Inc. | PD-L1-binding molecules comprising Shiga toxin a subunit scaffolds |
BR112022004972A2 (en) | 2019-09-18 | 2022-06-28 | Genentech Inc | ANTIBODIES, ISOLATED NUCLEIC ACIDS, ISOLATED HOST CELLS, METHODS OF PRODUCTION OF AN ANTIBODY, OF PRODUCTION OF A BIESPECIFIC ANTIBODY, AND OF TREATMENT OF AN INDIVIDUAL, BIESPECIFIC ANTIBODIES, PHARMACEUTICAL COMPOSITION, ANTIBODY, BIESPECIFIC ANTIBODY OR PHARMACEUTICAL COMPOSITION OF THE ANTIBODY, USE OF A COMBINATION OF THE ANTIBODY AND METHODS TO REDUCE INFLAMMATION AND TO IMPROVE SCALING AND/OR SKIN ERUPTION AND COMBINATION FOR USE |
JP2022549218A (en) | 2019-09-20 | 2022-11-24 | ジェネンテック, インコーポレイテッド | Anti-tryptase antibody medication |
CN112625130B (en) | 2019-09-24 | 2023-08-29 | 财团法人工业技术研究院 | Anti-TIGIT antibodies and methods of use |
JP2022550325A (en) | 2019-09-27 | 2022-12-01 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Anti-Mullerian inhibitor antibody and use thereof |
WO2021057978A1 (en) | 2019-09-27 | 2021-04-01 | 南京金斯瑞生物科技有限公司 | Anti-vhh domain antibodies and use thereof |
WO2021058729A1 (en) | 2019-09-27 | 2021-04-01 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Anti-müllerian inhibiting substance type i receptor antibodies and uses thereof |
WO2021062085A1 (en) | 2019-09-27 | 2021-04-01 | Genentech, Inc. | Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies |
CA3151450A1 (en) | 2019-09-30 | 2021-04-08 | Matthias Schneider | Protein binders to irhom2 epitopes |
WO2021063968A1 (en) | 2019-09-30 | 2021-04-08 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Method and composition for diagnosing chronic obstructive pulmonary disease |
TW202128756A (en) | 2019-10-02 | 2021-08-01 | 德商百靈佳殷格翰國際股份有限公司 | Multi-specific binding proteins for cancer treatment |
WO2021064180A1 (en) | 2019-10-03 | 2021-04-08 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for modulating macrophages polarization |
US20220363776A1 (en) | 2019-10-04 | 2022-11-17 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical composition for the treatment of ovarian cancer, breast cancer or pancreatic cancer |
BR112022007216A2 (en) | 2019-10-18 | 2022-08-23 | Genentech Inc | METHODS FOR TREATMENT OF DIFFUSE LYMPHOMA, KIT AND IMMUNOCONJUGATE |
WO2021078359A1 (en) | 2019-10-21 | 2021-04-29 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of inhibitors of cubilin for the treatment of chronic kidney diseases |
WO2021089513A1 (en) | 2019-11-05 | 2021-05-14 | F. Hoffmann-La Roche Ag | Treatment of cancer using a hla-a2/wt1 x cd3 bispecific antibody and lenalidomide |
JP2022553803A (en) | 2019-11-06 | 2022-12-26 | ジェネンテック, インコーポレイテッド | Diagnostic and therapeutic methods for the treatment of blood cancers |
CA3159541A1 (en) | 2019-11-07 | 2021-05-14 | Eisai R&D Management Co., Ltd. | Anti-mesothelin eribulin antibody-drug conjugates and methods of use |
JP2023500954A (en) | 2019-11-12 | 2023-01-11 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | A new serum marker for the latent form of toxoplasmosis |
KR20220113386A (en) | 2019-11-15 | 2022-08-12 | 플라이언트 테라퓨틱스, 인크. | Compositions and methods for activation of integrins |
WO2021110796A1 (en) | 2019-12-04 | 2021-06-10 | Bayer Aktiengesellschaft | Inhibitors of shp2 |
EP4072682A1 (en) | 2019-12-09 | 2022-10-19 | Institut National de la Santé et de la Recherche Médicale (INSERM) | Antibodies having specificity to her4 and uses thereof |
GB201918279D0 (en) | 2019-12-12 | 2020-01-29 | Vib Vzw | Glycosylated single chain immunoglobulin domains |
MX2022007158A (en) | 2019-12-13 | 2022-07-11 | Genentech Inc | Anti-ly6g6d antibodies and methods of use. |
IL293827A (en) | 2019-12-13 | 2022-08-01 | Alector Llc | Anti-mertk antibodies and methods of use thereof |
WO2021127217A1 (en) | 2019-12-17 | 2021-06-24 | Flagship Pioneering Innovations V, Inc. | Combination anti-cancer therapies with inducers of iron-dependent cellular disassembly |
AU2020406085A1 (en) | 2019-12-18 | 2022-05-26 | F. Hoffmann-La Roche Ag | Antibodies binding to HLA-A2/MAGE-A4 |
AU2020408213A1 (en) | 2019-12-19 | 2022-06-23 | Quidel Corporation | Monoclonal antibody fusions |
BR112022012439A2 (en) | 2019-12-23 | 2022-09-06 | Genentech Inc | ISOLATED ANTIBODIES, ISOLATED NUCLEIC ACID, ISOLATED VECTOR, ISOLATED HOST CELL, AND METHODS TO PRODUCE AN ANTIBODY THAT BINDS TO APOL1, TO DETECT APOLIPOPROTEIN L1 IN A SAMPLE, TO DISTINGUISH ENDOGENOUS APOLIPOPROTEIN L1, TO DISTINGUISH THE G0 AND G1 FORMS OF APOLIPOPROTEIN L1 APOL1 G2 E FORM TO SPECIFICALLY DETECT PODOCYTE CELLS |
MX2022007798A (en) | 2019-12-23 | 2022-07-19 | Eisai R&D Man Co Ltd | Method for producing eribulin-based antibody-drug conjugate. |
MX2022007840A (en) | 2019-12-27 | 2022-07-19 | Chugai Pharmaceutical Co Ltd | Anti-ctla-4 antibody and use thereof. |
CN114929734A (en) | 2020-01-09 | 2022-08-19 | 豪夫迈·罗氏有限公司 | Novel antigen binding molecules comprising 4-1BBL trimers |
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 |
WO2021144426A1 (en) | 2020-01-17 | 2021-07-22 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
WO2021194481A1 (en) | 2020-03-24 | 2021-09-30 | Genentech, Inc. | Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies |
WO2022050954A1 (en) | 2020-09-04 | 2022-03-10 | Genentech, Inc. | Dosing for treatment with anti-tigit and anti-pd-l1 antagonist antibodies |
CN115397459A (en) | 2020-01-31 | 2022-11-25 | 基因泰克公司 | Method for inducing new epitope-specific T cells using PD-1 axis binding antagonists and RNA vaccines |
US20230061973A1 (en) | 2020-02-05 | 2023-03-02 | Larimar Therapeutics, Inc. | Tat peptide binding proteins and uses thereof |
KR20220139357A (en) | 2020-02-10 | 2022-10-14 | 상하이 에스쿠겐 바이오테크놀로지 컴퍼니 리미티드 | CLDN18.2 Antibodies and Their Uses |
JP2023513401A (en) | 2020-02-10 | 2023-03-30 | 上海詩健生物科技有限公司 | Antibodies to claudin 18.2 and uses thereof |
TW202144395A (en) | 2020-02-12 | 2021-12-01 | 日商中外製藥股份有限公司 | Anti-CD137 antigen-binding molecule for use in cancer treatment |
CN113248611A (en) | 2020-02-13 | 2021-08-13 | 湖南华康恒健生物技术有限公司 | anti-BCMA antibody, pharmaceutical composition and application thereof |
JP2023514957A (en) | 2020-02-28 | 2023-04-12 | オレガ・バイオテック | Combination therapy based on CTLA4 inhibitors and IL-17B inhibitors |
WO2021183849A1 (en) | 2020-03-13 | 2021-09-16 | Genentech, Inc. | Anti-interleukin-33 antibodies and uses thereof |
CR20220524A (en) | 2020-03-19 | 2022-12-02 | Genentech Inc | Isoform-selective anti-tgf-beta antibodies and methods of use |
MX2022011752A (en) | 2020-03-24 | 2022-10-18 | Genentech Inc | Tie2-binding agents and methods of use. |
JP2023518841A (en) | 2020-03-26 | 2023-05-08 | ジェネンテック, インコーポレイテッド | Modified mammalian cells with reduced host cell proteins |
WO2021202590A1 (en) | 2020-03-31 | 2021-10-07 | Alector Llc | Anti-mertk antibodies and methods of use thereof |
CN115335410A (en) | 2020-03-31 | 2022-11-11 | 中外制药株式会社 | Method for producing multispecific antigen-binding molecules |
CN115915939A (en) | 2020-03-31 | 2023-04-04 | 生物催化公司 | Antifungal polypeptides |
AR121706A1 (en) | 2020-04-01 | 2022-06-29 | Hoffmann La Roche | OX40 AND FAP-TARGETED BSPECIFIC ANTIGEN-BINDING MOLECULES |
JP2023519930A (en) | 2020-04-01 | 2023-05-15 | ユニバーシティ オブ ロチェスター | Monoclonal Antibodies Against Hemagglutinin (HA) and Neuraminidase (NA) of Influenza H3N2 Virus |
EP4127724A1 (en) | 2020-04-03 | 2023-02-08 | Genentech, Inc. | Therapeutic and diagnostic methods for cancer |
WO2021198511A1 (en) | 2020-04-03 | 2021-10-07 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treatment of sars-cov-2 infection |
US20230192795A1 (en) | 2020-04-15 | 2023-06-22 | Hoffmann-La Roche Inc. | Immunoconjugates |
EP4144758A1 (en) | 2020-04-22 | 2023-03-08 | Mabwell (Shanghai) Bioscience Co., Ltd. | Single variable domain antibody targeting human programmed death ligand 1 (pd-l1) and derivative thereof |
IL297541A (en) | 2020-04-24 | 2022-12-01 | Genentech Inc | Methods of using anti-cd79b immunoconjugates |
IL296664A (en) | 2020-04-24 | 2022-11-01 | Hoffmann La Roche | Enzyme and pathway modulation with sulfhydryl compounds and their derivatives |
TW202206100A (en) | 2020-04-27 | 2022-02-16 | 美商西健公司 | Treatment for cancer |
US11634477B2 (en) | 2020-04-28 | 2023-04-25 | The Rockefeller University | Neutralizing anti-SARS-CoV-2 antibodies and methods of use thereof |
CN115885050A (en) | 2020-04-28 | 2023-03-31 | 基因泰克公司 | 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 |
WO2021224401A1 (en) | 2020-05-07 | 2021-11-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for determining a reference range of β-galactose exposure platelet |
CA3180683A1 (en) | 2020-05-12 | 2021-11-18 | Inserm (Institut National De La Sante Et De La Recherche Medicale) | New method to treat cutaneous t-cell lymphomas and tfh derived lymphomas |
WO2021229104A1 (en) | 2020-05-15 | 2021-11-18 | Université de Liège | Anti-cd38 single-domain antibodies in disease monitoring and treatment |
US20230212260A1 (en) | 2020-05-15 | 2023-07-06 | Apogenix Ag | Multi-specific immune modulators |
US20210363273A1 (en) | 2020-05-19 | 2021-11-25 | Boehringer Ingelheim International Gmbh | Binding molecules for the treatment of cancer |
WO2021236845A1 (en) | 2020-05-20 | 2021-11-25 | Takeda Vaccines, Inc. | Method for detection of zika virus specific antibodies |
JP2023526477A (en) | 2020-05-20 | 2023-06-21 | アンスティテュ・クリー | Synthetic single domain library |
JP2023527169A (en) | 2020-05-20 | 2023-06-27 | タケダ ワクチン,インコーポレイテッド | Methods for Determining Antigen Potency |
WO2021236225A1 (en) | 2020-05-20 | 2021-11-25 | Takeda Vaccines, Inc. | Method for detection of zika virus specific antibodies |
JP2023528293A (en) | 2020-05-20 | 2023-07-04 | アンスティテュ・クリー | Single domain antibodies and their use in cancer therapy |
BR112022024339A2 (en) | 2020-05-29 | 2022-12-27 | 23Andme Inc | ANTI CD200R1 ANTIBODIES AND METHODS OF THEIR USE |
CA3184645A1 (en) | 2020-06-05 | 2021-12-09 | Eisai R&D Management Co., Ltd. | Anti-bcma antibody-drug conjugates and methods of use |
JP2023527578A (en) | 2020-06-05 | 2023-06-29 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Methods and pharmaceutical compositions for treating eye diseases |
CA3184495A1 (en) | 2020-06-08 | 2021-12-16 | F. Hoffmann-La Roche Ag | Anti-hbv antibodies and methods of use |
CN115803063A (en) | 2020-06-11 | 2023-03-14 | 基因泰克公司 | Nanolipid protein-polypeptide conjugates and compositions, systems, and methods of use thereof |
CN115698719A (en) | 2020-06-12 | 2023-02-03 | 基因泰克公司 | Methods and compositions for cancer immunotherapy |
CN115916182A (en) | 2020-06-16 | 2023-04-04 | 基因泰克公司 | 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 |
CA3185513A1 (en) | 2020-06-19 | 2021-12-23 | F. Hoffmann-La Roche Ag | Antibodies binding to cd3 and folr1 |
MX2022016069A (en) | 2020-06-19 | 2023-02-02 | Hoffmann La Roche | Antibodies binding to cd3 and cd19. |
WO2021255146A1 (en) | 2020-06-19 | 2021-12-23 | F. Hoffmann-La Roche Ag | Antibodies binding to cd3 and cea |
AU2021291002A1 (en) | 2020-06-19 | 2022-10-13 | F. Hoffmann-La Roche Ag | Protease-activated T cell bispecific antibodies |
JP2023530961A (en) | 2020-06-19 | 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 | Antibody that binds to CD3 |
WO2021259880A1 (en) | 2020-06-22 | 2021-12-30 | Almirall, S.A. | Anti-il-36 antibodies and methods of use thereof |
PE20231361A1 (en) | 2020-06-23 | 2023-09-05 | Hoffmann La Roche | AGONIST MOLECULES BINDING TO THE CD28 ANTIGEN THAT TARGETS HER2 |
US20220041672A1 (en) | 2020-06-24 | 2022-02-10 | Genentech, Inc. | Apoptosis resistant cell lines |
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 |
US20230355804A1 (en) | 2020-06-29 | 2023-11-09 | Flagship Pioneering Innovations V, Inc. | Viruses engineered to promote thanotransmission and their use in treating cancer |
CN115884986A (en) | 2020-06-29 | 2023-03-31 | Inserm(法国国家健康医学研究院) | Anti-protein S single domain antibodies and polypeptides comprising the same |
WO2022008597A1 (en) | 2020-07-08 | 2022-01-13 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical composition for the treatment of infectious diseases |
WO2022015726A1 (en) | 2020-07-13 | 2022-01-20 | Genentech, Inc. | Cell-based methods for predicting polypeptide immunogenicity |
JP2023534458A (en) | 2020-07-17 | 2023-08-09 | ジェネンテック, インコーポレイテッド | Anti-Notch2 antibody and method of use |
KR20230042032A (en) | 2020-07-21 | 2023-03-27 | 제넨테크, 인크. | Antibody Conjugation Chemical Inducers of BRM Degradation and Methods Thereof |
GB2597532A (en) | 2020-07-28 | 2022-02-02 | Femtogenix Ltd | Cytotoxic compounds |
WO2022023379A1 (en) | 2020-07-28 | 2022-02-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for preventing and treating a cancer |
WO2022029080A1 (en) | 2020-08-03 | 2022-02-10 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Population of treg cells functionally committed to exert a regulatory activity and their use for adoptive therapy |
JP2023537683A (en) | 2020-08-07 | 2023-09-05 | ジェネンテック, インコーポレイテッド | T cell-based methods for predicting polypeptide immunogenicity |
CA3187245A1 (en) | 2020-08-17 | 2022-02-24 | Sebastien Mercx | Recombinant immunotoxin comprising a ribotoxin or rnase |
CN114106173A (en) | 2020-08-26 | 2022-03-01 | 上海泰槿生物技术有限公司 | anti-OX 40 antibodies, pharmaceutical compositions and uses thereof |
TW202227625A (en) | 2020-08-28 | 2022-07-16 | 美商建南德克公司 | Crispr/cas9 multiplex knockout of host cell proteins |
WO2022047381A1 (en) | 2020-08-31 | 2022-03-03 | Genentech, Inc. | Methods for producing antibodies |
EP4211165A1 (en) | 2020-09-14 | 2023-07-19 | Ichnos Sciences SA | Antibodies that bind to il1rap and uses thereof |
WO2022063957A1 (en) | 2020-09-24 | 2022-03-31 | Vib Vzw | Biomarker for anti-tumor therapy |
AU2021350075A1 (en) | 2020-09-24 | 2023-03-09 | F. Hoffmann-La Roche Ag | Prevention or mitigation of T-cell bispecific antibody-related adverse effects |
US20230364049A1 (en) | 2020-09-24 | 2023-11-16 | Vib Vzw | Combination of p2y6 inhibitors and immune checkpoint inhibitors |
TW202229346A (en) | 2020-09-28 | 2022-08-01 | 美商西根公司 | Humanized anti-liv1 antibodies for the treatment of cancer |
WO2022064049A1 (en) | 2020-09-28 | 2022-03-31 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Method for diagnosing brucella infection |
TW202229348A (en) | 2020-09-30 | 2022-08-01 | 美商西健公司 | Uveal melanoma treatment using sea-cd40 |
CN116406291A (en) | 2020-10-05 | 2023-07-07 | 基因泰克公司 | Administration of treatment with anti-FCRH 5/anti-CD 3 bispecific antibodies |
WO2022076865A1 (en) | 2020-10-09 | 2022-04-14 | Takeda Vaccines, Inc. | Methods for determining complement-fixing antibodies |
EP4229090A1 (en) | 2020-10-16 | 2023-08-23 | Université d'Aix-Marseille | Anti-gpc4 single domain antibodies |
WO2022084300A1 (en) | 2020-10-20 | 2022-04-28 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for diagnosis and monitoring form of coronavirus infection |
CA3190782A1 (en) | 2020-10-20 | 2022-04-28 | F. Hoffmann-La Roche Ag | Combination therapy of pd-1 axis binding antagonists and lrrk2 inhibitors |
WO2022086957A1 (en) | 2020-10-20 | 2022-04-28 | Genentech, Inc. | Peg-conjugated anti-mertk antibodies and methods of use |
JP2023546229A (en) | 2020-10-21 | 2023-11-01 | ベーリンガー インゲルハイム インターナショナル ゲゼルシャフト ミット ベシュレンクテル ハフツング | Agonistic TrkB binding molecules for the treatment of ocular diseases |
WO2022084531A1 (en) | 2020-10-23 | 2022-04-28 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating glioma |
WO2022093981A1 (en) | 2020-10-28 | 2022-05-05 | Genentech, Inc. | Combination therapy comprising ptpn22 inhibitors and pd-l1 binding antagonists |
JP2023549062A (en) | 2020-10-30 | 2023-11-22 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Treatment of cancer using CEA CD3 bispecific antibodies and TGFβ signaling inhibitors |
CA3196076A1 (en) | 2020-11-04 | 2022-05-12 | Chi-Chung Li | Subcutaneous dosing of anti-cd20/anti-cd3 bispecific antibodies |
IL302639A (en) | 2020-11-04 | 2023-07-01 | Myeloid Therapeutics Inc | Engineered chimeric fusion protein compositions and methods of use thereof |
CA3196191A1 (en) | 2020-11-04 | 2022-05-12 | Chi-Chung Li | Dosing for treatment with anti-cd20/anti-cd3 bispecific antibodies and anti-cd79b antibody drug conjugates |
KR20230095119A (en) | 2020-11-04 | 2023-06-28 | 제넨테크, 인크. | Dosing for Treatment with Anti-CD20/Anti-CD3 Bispecific Antibodies |
EP4240758A1 (en) | 2020-11-04 | 2023-09-13 | The Rockefeller University | Neutralizing anti-sars-cov-2 antibodies |
WO2022096547A1 (en) | 2020-11-05 | 2022-05-12 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of il-6 inhibitors for the treatment of acute chest syndrome in patients suffering from sickle cell disease |
KR20230107235A (en) | 2020-11-10 | 2023-07-14 | 에프. 호프만-라 로슈 아게 | Prevention or alleviation of negative effects associated with T-cell engagement agents |
PE20231556A1 (en) | 2020-11-16 | 2023-10-03 | Hoffmann La Roche | GLYCOFORMS FROM MANNOSE FACTORIES |
WO2022101481A1 (en) | 2020-11-16 | 2022-05-19 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for predicting and treating uveal melanoma |
WO2022101458A1 (en) | 2020-11-16 | 2022-05-19 | F. Hoffmann-La Roche Ag | Combination therapy with fap-targeted cd40 agonists |
WO2022109441A1 (en) | 2020-11-23 | 2022-05-27 | Genentech, Inc. | Methods for modulating host cell surface interactions with sars-cov-2 |
AU2021386240A1 (en) | 2020-11-27 | 2023-06-29 | Genentech, Inc. | Methods for modulating host cell surface interactions with human cytomegalovirus |
JP2023552748A (en) | 2020-12-01 | 2023-12-19 | ジェネンテック, インコーポレイテッド | Biological vesicles displaying cell surface proteins and related methods |
US20240101681A1 (en) | 2020-12-02 | 2024-03-28 | Alector Llc | Methods of use of anti-sortilin antibodies |
IL303656A (en) | 2020-12-17 | 2023-08-01 | Hoffmann La Roche | Anti-hla-g antibodies and use thereof |
CN117015386A (en) | 2020-12-21 | 2023-11-07 | 旗舰创业创新五公司 | Use of cell turnover factors for increasing tissue regeneration |
AU2022206061A1 (en) | 2021-01-06 | 2023-07-06 | F. Hoffmann-La Roche Ag | Combination therapy employing a pd1-lag3 bispecific antibody and a cd20 t cell bispecific antibody |
US20240115720A1 (en) | 2021-01-13 | 2024-04-11 | Memorial Sloan Kettering Cancer Center | Antibody-pyrrolobenzodiazepine derivative conjugate |
WO2022153212A1 (en) | 2021-01-13 | 2022-07-21 | Axon Neuroscience Se | Antibodies neutralizing sars-cov-2 |
JP2024503658A (en) | 2021-01-13 | 2024-01-26 | メモリアル スローン-ケタリング キャンサー センター | Anti-DLL3 antibody-drug conjugate |
JP2024505428A (en) | 2021-01-14 | 2024-02-06 | アンスティテュ キュリー | HER2 single domain antibody variants and their CARs |
JP2024505636A (en) | 2021-01-15 | 2024-02-07 | ザ ロックフェラー ユニバーシティー | Anti-SARS-COV-2 neutralizing antibody |
AR124681A1 (en) | 2021-01-20 | 2023-04-26 | Abbvie Inc | ANTI-EGFR ANTIBODY-DRUG CONJUGATES |
CN116917326A (en) | 2021-01-22 | 2023-10-20 | 博泰康医药公司 | anti-HER-2/TROP-2 constructs and uses thereof |
JP2024504475A (en) | 2021-01-28 | 2024-01-31 | 南京▲樺▼冠生物技▲術▼有限公司 | Complexes and their use |
CN117241804A (en) | 2021-02-17 | 2023-12-15 | 非营利性组织佛兰芒综合大学生物技术研究所 | Inhibition of SLC4A4 in cancer treatment |
CN117321076A (en) | 2021-02-19 | 2023-12-29 | 美国卫生及公众服务部代表 | Single domain antibodies neutralizing SARS-CoV-2 |
CA3207134A1 (en) | 2021-02-19 | 2022-08-25 | Jeffrey A. Ledbetter | Dnase fusion polypeptides and related compositions and methods |
TW202241941A (en) | 2021-02-22 | 2022-11-01 | 美商建南德克公司 | Methods for modulating host cell surface interactions with herpesviruses |
EP4298124A1 (en) | 2021-02-26 | 2024-01-03 | Bayer Aktiengesellschaft | Inhibitors of il-11 or il-11ra for use in the treatment of abnormal uterine bleeding |
TW202246324A (en) | 2021-03-01 | 2022-12-01 | 美商艾希利歐發展股份有限公司 | Combination of masked ctla4 and pd1/pdl1 antibodies for treating cancer |
TW202317612A (en) | 2021-03-01 | 2023-05-01 | 美商艾希利歐發展股份有限公司 | Combination of ctla4 and pd1/pdl1 antibodies for treating cancer |
JP2024510415A (en) | 2021-03-01 | 2024-03-07 | スキロム ゲゼルシャフト ミット ベシュレンクテル ハフツング | Humanized antibody against iRhom2 |
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 |
KR20230156387A (en) | 2021-03-12 | 2023-11-14 | 얀센 바이오테크 인코포레이티드 | Safe and effective method of treating psoriatic arthritis by anti-IL23 specific antibody |
AR125074A1 (en) | 2021-03-12 | 2023-06-07 | Genentech Inc | ANTI-KLK7 ANTIBODIES, ANTI-KLK5 ANTIBODIES, ANTI-KLK5/KLK7 MULTI-SPECIFIC ANTIBODIES AND METHODS OF USE |
CA3213278A1 (en) | 2021-03-12 | 2022-09-15 | Janssen Biotech, Inc. | Method of treating psoriatic arthritis patients with inadequate response to tnf therapy with anti-il23 specific antibody |
WO2022198192A1 (en) | 2021-03-15 | 2022-09-22 | Genentech, Inc. | Compositions and methods of treating lupus nephritis |
IL305901A (en) | 2021-03-17 | 2023-11-01 | Receptos Llc | Methods of treating atopic dermatitis with anti il-13 antibodies |
WO2022197945A1 (en) | 2021-03-17 | 2022-09-22 | Molecular Templates, Inc. | Pd-l1 binding proteins comprising shiga toxin a subunit scaffolds and cd8+ t cell antigens |
EP4308118A1 (en) | 2021-03-17 | 2024-01-24 | Institut National de la Santé et de la Recherche Médicale (INSERM) | Methods and compositions for treating melanoma |
EP4308606A1 (en) | 2021-03-18 | 2024-01-24 | Alector LLC | Anti-tmem106b antibodies and methods of use thereof |
WO2022197877A1 (en) | 2021-03-19 | 2022-09-22 | Genentech, Inc. | Methods and compositions for time delayed bio-orthogonal release of cytotoxic agents |
WO2022200303A1 (en) | 2021-03-23 | 2022-09-29 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for the diagnosis and treatment of t cell-lymphomas |
EP4314063A1 (en) | 2021-03-23 | 2024-02-07 | Alector LLC | Anti-tmem106b antibodies for treating and preventing coronavirus infections |
CA3213771A1 (en) | 2021-03-29 | 2022-10-06 | Scirhom Gmbh | Methods of treatment using protein binders to irhom2 epitopes |
CN117157312A (en) | 2021-03-30 | 2023-12-01 | 豪夫迈·罗氏有限公司 | Protease-activated polypeptides |
WO2022212784A1 (en) | 2021-03-31 | 2022-10-06 | Flagship Pioneering Innovations V, Inc. | Thanotransmission polypeptides and their use in treating cancer |
JP2024515591A (en) | 2021-04-08 | 2024-04-10 | マレンゴ・セラピューティクス,インコーポレーテッド | Multifunctional molecules that bind to TCRs and uses thereof |
EP4320153A1 (en) | 2021-04-09 | 2024-02-14 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for the treatment of anaplastic large cell lymphoma |
EP4323402A1 (en) | 2021-04-14 | 2024-02-21 | Villaris Therapeutics, Inc. | Anti-cd122 antibodies and uses thereof |
AR125344A1 (en) | 2021-04-15 | 2023-07-05 | Chugai Pharmaceutical Co Ltd | ANTI-C1S ANTIBODY |
TW202305122A (en) | 2021-04-19 | 2023-02-01 | 美商建南德克公司 | Modified mammalian cells |
EP4326871A1 (en) | 2021-04-19 | 2024-02-28 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of splice switching oligonucleotides for exon skipping-mediated knockdown of pim2 |
WO2022223791A1 (en) | 2021-04-23 | 2022-10-27 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating cell senescence accumulation related disease |
WO2022223651A1 (en) | 2021-04-23 | 2022-10-27 | F. Hoffmann-La Roche Ag | Prevention or mitigation of nk cell engaging agent-related adverse effects |
AU2021443863A1 (en) | 2021-04-30 | 2023-10-26 | F. Hoffmann-La Roche Ag | Dosing for treatment with anti-cd20/anti-cd3 bispecific antibody |
KR20240005691A (en) | 2021-04-30 | 2024-01-12 | 에프. 호프만-라 로슈 아게 | Dosage for combination therapy with anti-CD20/anti-CD3 bispecific antibody and anti-CD79B antibody drug conjugate |
WO2022235867A2 (en) | 2021-05-06 | 2022-11-10 | The Rockefeller University | Neutralizing anti-sars- cov-2 antibodies and methods of use thereof |
BR112023023622A2 (en) | 2021-05-12 | 2024-02-06 | Genentech Inc | METHODS TO TREAT DIFFUSE LYMPHOMA, KITS, IMMUNOCONJUGATES, POLATUZUMABE VEDOTIN AND IMMUNOCONJUGATE FOR USE |
AR125855A1 (en) | 2021-05-14 | 2023-08-16 | Genentech Inc | TREM2 AGONISTS |
WO2022242892A1 (en) | 2021-05-17 | 2022-11-24 | Université de Liège | Anti-cd38 single-domain antibodies in disease monitoring and treatment |
WO2022243261A1 (en) | 2021-05-19 | 2022-11-24 | F. Hoffmann-La Roche Ag | Agonistic cd40 antigen binding molecules targeting cea |
KR20240010469A (en) | 2021-05-21 | 2024-01-23 | 제넨테크, 인크. | Modified cells for production of recombinant products of interest |
TW202314247A (en) | 2021-05-25 | 2023-04-01 | 美商思進公司 | Methods of quantifying anti-cd40 antibodies |
TW202306979A (en) | 2021-05-26 | 2023-02-16 | 美商建南德克公司 | Methods for modulating host cell surface interactions with human cytomegalovirus |
AR126009A1 (en) | 2021-06-02 | 2023-08-30 | Hoffmann La Roche | CD28 ANTIGEN-BINDING AGONIST MOLECULES THAT TARGET EPCAM |
CN117480184A (en) | 2021-06-04 | 2024-01-30 | 中外制药株式会社 | anti-DDR 2 antibodies and uses thereof |
US11572412B2 (en) | 2021-06-04 | 2023-02-07 | Boehringer Ingelheim International Gmbh | Anti-SIRP-alpha antibodies |
WO2022258600A1 (en) | 2021-06-09 | 2022-12-15 | F. Hoffmann-La Roche Ag | Combination of a particular braf inhibitor (paradox breaker) and a pd-1 axis binding antagonist for use in the treatment of cancer |
WO2022266221A1 (en) | 2021-06-16 | 2022-12-22 | Alector Llc | Monovalent anti-mertk antibodies and methods of use thereof |
WO2022266223A1 (en) | 2021-06-16 | 2022-12-22 | Alector Llc | Bispecific anti-mertk and anti-pdl1 antibodies and methods of use thereof |
AR126161A1 (en) | 2021-06-17 | 2023-09-27 | Boehringer Lngelheim Int Gmbh | NOVEL TRISPECIFIC BINDING MOLECULES |
WO2022269473A1 (en) | 2021-06-23 | 2022-12-29 | Janssen Biotech, Inc. | Materials and methods for hinge regions in functional exogenous receptors |
IL308633A (en) | 2021-06-25 | 2024-01-01 | Chugai Pharmaceutical Co Ltd | Use of anti-ctla-4 antibody |
CA3221833A1 (en) | 2021-06-25 | 2022-12-29 | Chugai Seiyaku Kabushiki Kaisha | Anti-ctla-4 antibody |
CA3224374A1 (en) | 2021-06-29 | 2023-01-05 | Flagship Pioneering Innovations V, Inc. | Immune cells engineered to promote thanotransmission and uses thereof |
TW202309078A (en) | 2021-07-02 | 2023-03-01 | 美商建南德克公司 | Methods and compositions for treating cancer |
WO2023281463A1 (en) | 2021-07-09 | 2023-01-12 | Janssen Biotech, Inc. | Manufacturing methods for producing anti-tnf antibody compositions |
IL309987A (en) | 2021-07-09 | 2024-03-01 | Janssen Biotech Inc | Manufacturing methods for producing anti-il12/il23 antibody compositions |
IL309997A (en) | 2021-07-09 | 2024-03-01 | Janssen Biotech Inc | Manufacturing methods for producing anti-tnf antibody compositions |
IL309559A (en) | 2021-07-09 | 2024-02-01 | Luxembourg Inst Of Health Lih | Dimeric protein complexes and uses thereof |
WO2023285362A1 (en) | 2021-07-12 | 2023-01-19 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Use of il-36 inhibitors for the treatment of netherton syndrome |
US20230049152A1 (en) | 2021-07-14 | 2023-02-16 | Genentech, Inc. | Anti-c-c motif chemokine receptor 8 (ccr8) antibodies and methods of use |
KR20240036570A (en) | 2021-07-22 | 2024-03-20 | 에프. 호프만-라 로슈 아게 | Heterodimeric Fc domain antibodies |
WO2023004386A1 (en) | 2021-07-22 | 2023-01-26 | Genentech, Inc. | Brain targeting compositions and methods of use thereof |
CA3224180A1 (en) | 2021-07-28 | 2023-02-02 | F. Hoffmann-La Roche Ag | Methods and compositions for treating cancer |
WO2023006975A2 (en) | 2021-07-30 | 2023-02-02 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Chimeric proteins and methods of immunotherapy |
WO2023010080A1 (en) | 2021-07-30 | 2023-02-02 | Seagen Inc. | Treatment for cancer |
WO2023012147A1 (en) | 2021-08-03 | 2023-02-09 | F. Hoffmann-La Roche Ag | Bispecific antibodies and methods of use |
WO2023019239A1 (en) | 2021-08-13 | 2023-02-16 | Genentech, Inc. | Dosing for anti-tryptase antibodies |
WO2023028591A1 (en) | 2021-08-27 | 2023-03-02 | Genentech, Inc. | Methods of treating tau pathologies |
TW202325727A (en) | 2021-08-30 | 2023-07-01 | 美商建南德克公司 | Anti-polyubiquitin multispecific antibodies |
TW202313695A (en) | 2021-09-15 | 2023-04-01 | 法商感應檢查療法公司 | Use of anti-btn3a antibody in manufacturing a medicament for use in treating a tumor |
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 |
WO2023056362A1 (en) | 2021-09-30 | 2023-04-06 | Seagen Inc. | B7-h4 antibody-drug conjugates for the treatment of cancer |
WO2023052541A1 (en) | 2021-09-30 | 2023-04-06 | Imcheck Therapeutics | Combination of an anti-btn3a activating antibody and an il-2 agonist for use in therapy |
WO2023060086A1 (en) | 2021-10-04 | 2023-04-13 | Takeda Vaccines, Inc. | Methods for determining norovirus-reactive antibodies |
WO2023057601A1 (en) | 2021-10-06 | 2023-04-13 | Biotalys NV | Anti-fungal polypeptides |
US20230190807A1 (en) | 2021-10-06 | 2023-06-22 | Immatics Biotechnologies Gmbh | Tcr compounds, compositions, and methods of treating |
WO2023062048A1 (en) | 2021-10-14 | 2023-04-20 | F. Hoffmann-La Roche Ag | Alternative pd1-il7v immunoconjugates for the treatment of cancer |
AU2022362681A1 (en) | 2021-10-14 | 2024-04-04 | F. Hoffmann-La Roche Ag | New interleukin-7 immunoconjugates |
WO2023069919A1 (en) | 2021-10-19 | 2023-04-27 | Alector Llc | Anti-cd300lb antibodies and methods of use thereof |
WO2023073084A1 (en) | 2021-10-27 | 2023-05-04 | Imcheck Therapeutics | Butyrophilin (btn) 3a activating antibodies for use in methods for treating infectious disorders |
WO2023073615A1 (en) | 2021-10-29 | 2023-05-04 | Janssen Biotech, Inc. | Methods of treating crohn's disease with anti-il23 specific antibody |
WO2023073225A1 (en) | 2021-11-01 | 2023-05-04 | F. Hoffmann-La Roche Ag | Treatment of cancer using a hla-a2/wt1 x cd3 bispecific antibody and a 4-1bb (cd137) agonist |
WO2023078900A1 (en) | 2021-11-03 | 2023-05-11 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating triple negative breast cancer (tnbc) |
WO2023081925A1 (en) | 2021-11-08 | 2023-05-11 | Immatics Biotechnologies Gmbh | Adoptive cell therapy combination treatment and compositions thereof |
US20230272082A1 (en) | 2021-11-09 | 2023-08-31 | Sensei Biotherapeutics, Inc. | Anti-vista antibodies and uses thereof |
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 |
WO2023088889A1 (en) | 2021-11-16 | 2023-05-25 | Apogenix Ag | CD137 ligands |
WO2023091887A1 (en) | 2021-11-16 | 2023-05-25 | Genentech, Inc. | Methods and compositions for treating systemic lupus erythematosus (sle) with mosunetuzumab |
WO2023088876A1 (en) | 2021-11-16 | 2023-05-25 | Apogenix Ag | Multi-specific immune modulators |
US20230159633A1 (en) | 2021-11-23 | 2023-05-25 | Janssen Biotech, Inc. | Method of Treating Ulcerative Colitis with Anti-IL23 Specific Antibody |
WO2023094525A1 (en) | 2021-11-25 | 2023-06-01 | Veraxa Biotech Gmbh | Improved antibody-payload conjugates (apcs) prepared by site-specific conjugation utilizing genetic code expansion |
EP4186529A1 (en) | 2021-11-25 | 2023-05-31 | Veraxa Biotech GmbH | Improved antibody-payload conjugates (apcs) prepared by site-specific conjugation utilizing genetic code expansion |
WO2023094698A1 (en) | 2021-11-29 | 2023-06-01 | Ose Immunotherapeutics | Specific antagonist anti-sirpg antibodies |
AR127887A1 (en) | 2021-12-10 | 2024-03-06 | Hoffmann La Roche | ANTIBODIES THAT BIND CD3 AND PLAP |
TW202339797A (en) | 2021-12-14 | 2023-10-16 | 瑞士商赫孚孟拉羅股份公司 | Treatment of cancer using a hla-a2/mage-a4 x cd3 bispecific antibody and a 4-1bb (cd137) agonist |
WO2023110937A1 (en) | 2021-12-14 | 2023-06-22 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Depletion of nk cells for the treatment of adverse post-ischemic cardiac remodeling |
WO2023114544A1 (en) | 2021-12-17 | 2023-06-22 | Dana-Farber Cancer Institute, Inc. | Antibodies and uses thereof |
WO2023114543A2 (en) | 2021-12-17 | 2023-06-22 | Dana-Farber Cancer Institute, Inc. | Platform for antibody discovery |
WO2023117834A1 (en) | 2021-12-20 | 2023-06-29 | F. Hoffmann-La Roche Ag | Agonistic ltbr antibodies and bispecific antibodies comprising them |
WO2023118165A1 (en) | 2021-12-21 | 2023-06-29 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and compositions for treating melanoma |
EP4209508A1 (en) | 2022-01-11 | 2023-07-12 | Centre national de la recherche scientifique | Nanobodies for the deneddylating enzyme nedp1 |
US20230322958A1 (en) | 2022-01-19 | 2023-10-12 | Genentech, Inc. | Anti-Notch2 Antibodies and Conjugates and Methods of Use |
WO2023147328A1 (en) | 2022-01-26 | 2023-08-03 | Genentech, Inc. | Antibody-conjugated chemical inducers of degradation with hydolysable maleimide linkers and methods thereof |
WO2023147329A1 (en) | 2022-01-26 | 2023-08-03 | Genentech, Inc. | Antibody-conjugated chemical inducers of degradation and methods thereof |
WO2023147399A1 (en) | 2022-01-27 | 2023-08-03 | The Rockefeller University | Broadly neutralizing anti-sars-cov-2 antibodies targeting the n-terminal domain of the spike protein and methods of use thereof |
WO2023144235A1 (en) | 2022-01-27 | 2023-08-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for monitoring and treating warburg effect in patients with pi3k-related disorders |
WO2023144303A1 (en) | 2022-01-31 | 2023-08-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Cd38 as a biomarker and biotarget in t-cell lymphomas |
EP4231017A1 (en) | 2022-02-17 | 2023-08-23 | Promise Proteomics | Detection and quantification of anti-drug antibodies and anti-self antibodies |
WO2023156634A1 (en) | 2022-02-17 | 2023-08-24 | Atb Therapeutics | Recombinant immunotoxin comprising a ribosome inactivating protein |
WO2023173026A1 (en) | 2022-03-10 | 2023-09-14 | Sorrento Therapeutics, Inc. | Antibody-drug conjugates and uses thereof |
WO2023170247A1 (en) | 2022-03-11 | 2023-09-14 | Mablink Bioscience | Antibody-drug conjugates and their uses |
WO2023175171A1 (en) | 2022-03-18 | 2023-09-21 | Inserm (Institut National De La Sante Et De La Recherche Medicale) | Bk polyomavirus antibodies 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 |
WO2023186756A1 (en) | 2022-03-28 | 2023-10-05 | F. Hoffmann-La Roche Ag | Interferon gamma variants and antigen binding molecules comprising these |
US20230312703A1 (en) | 2022-03-30 | 2023-10-05 | Janssen Biotech, Inc. | Method of Treating Psoriasis with IL-23 Specific Antibody |
WO2023191816A1 (en) | 2022-04-01 | 2023-10-05 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
EP4257609A1 (en) | 2022-04-08 | 2023-10-11 | iOmx Therapeutics AG | Combination therapies based on pd-1 inhibitors and sik3 inhibitors |
WO2023198648A1 (en) | 2022-04-11 | 2023-10-19 | Institut National de la Santé et de la Recherche Médicale | Methods for the diagnosis and treatment of t-cell malignancies |
TW202400243A (en) | 2022-04-12 | 2024-01-01 | 日商衛材R&D企管股份有限公司 | Eribulin-based antibody-drug conjugates and methods of use |
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 |
US20230406930A1 (en) | 2022-04-13 | 2023-12-21 | Genentech, Inc. | Pharmaceutical compositions of therapeutic proteins and methods of use |
WO2023198851A1 (en) | 2022-04-14 | 2023-10-19 | Institut National de la Santé et de la Recherche Médicale | Methods for controlling the tumor cell killing by light |
WO2023198874A1 (en) | 2022-04-15 | 2023-10-19 | Institut National de la Santé et de la Recherche Médicale | Methods for the diagnosis and treatment of t cell-lymphomas |
WO2023212294A1 (en) | 2022-04-29 | 2023-11-02 | Broadwing Bio Llc | Angiopoietin-related protein 7-specific antibodies and uses thereof |
WO2023212298A1 (en) | 2022-04-29 | 2023-11-02 | Broadwing Bio Llc | Bispecific antibodies and methods of treating ocular disease |
WO2023212293A1 (en) | 2022-04-29 | 2023-11-02 | Broadwing Bio Llc | Complement factor h related 4-specific antibodies and uses thereof |
WO2023215737A1 (en) | 2022-05-03 | 2023-11-09 | Genentech, Inc. | Anti-ly6e antibodies, immunoconjugates, and uses thereof |
WO2023217904A1 (en) | 2022-05-10 | 2023-11-16 | Institut National de la Santé et de la Recherche Médicale | Syncitin-1 fusion proteins and uses thereof for cargo delivery into target cells |
WO2023219613A1 (en) | 2022-05-11 | 2023-11-16 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2023218431A1 (en) | 2022-05-13 | 2023-11-16 | BioNTech SE | Rna compositions targeting hiv |
WO2023223265A1 (en) | 2022-05-18 | 2023-11-23 | Janssen Biotech, Inc. | Method for evaluating and treating psoriatic arthritis with il23 antibody |
US20230416412A1 (en) | 2022-05-31 | 2023-12-28 | Hoffmann-La Roche Inc. | Prevention or mitigation of t-cell engaging agent-related adverse effects |
WO2023240058A2 (en) | 2022-06-07 | 2023-12-14 | Genentech, Inc. | Prognostic and therapeutic methods for cancer |
WO2023237661A1 (en) | 2022-06-09 | 2023-12-14 | Institut National de la Santé et de la Recherche Médicale | Use of endothelin receptor type b agonists for the treatment of aortic valve stenosis |
EP4299124A1 (en) | 2022-06-30 | 2024-01-03 | Universite De Montpellier | Anti-mglur2 nanobodies for use as biomolecule transporter |
WO2024003310A1 (en) | 2022-06-30 | 2024-01-04 | Institut National de la Santé et de la Recherche Médicale | Methods for the diagnosis and treatment of acute lymphoblastic leukemia |
WO2024008274A1 (en) | 2022-07-04 | 2024-01-11 | Universiteit Antwerpen | T regulatory cell modification |
WO2024008755A1 (en) | 2022-07-04 | 2024-01-11 | Vib Vzw | Blood-cerebrospinal fluid barrier crossing antibodies |
WO2024008799A1 (en) | 2022-07-06 | 2024-01-11 | Institut National de la Santé et de la Recherche Médicale | Methods for the treatment of proliferative glomerulonephritis |
WO2024015897A1 (en) | 2022-07-13 | 2024-01-18 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2024013234A1 (en) | 2022-07-13 | 2024-01-18 | Institut National de la Santé et de la Recherche Médicale | Methods for diagnosis, prognosis, stratification and treating of myocarditis |
US20240052065A1 (en) | 2022-07-15 | 2024-02-15 | Boehringer Ingelheim International Gmbh | Binding molecules for the treatment of cancer |
WO2024020432A1 (en) | 2022-07-19 | 2024-01-25 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2024018003A1 (en) | 2022-07-21 | 2024-01-25 | Institut National de la Santé et de la Recherche Médicale | Extracellular vesicles functionalized with an erv syncitin and uses thereof for cargo delivery |
WO2024018046A1 (en) | 2022-07-22 | 2024-01-25 | Institut National de la Santé et de la Recherche Médicale | Garp as a biomarker and biotarget in t-cell malignancies |
WO2024018426A1 (en) | 2022-07-22 | 2024-01-25 | Janssen Biotech, Inc. | Enhanced transfer of genetic instructions to effector immune cells |
WO2024020579A1 (en) | 2022-07-22 | 2024-01-25 | Bristol-Myers Squibb Company | Antibodies binding to human pad4 and uses thereof |
WO2024023246A1 (en) | 2022-07-28 | 2024-02-01 | Philogen S.P.A. | Antibody binding to pd1 |
WO2024026472A2 (en) | 2022-07-29 | 2024-02-01 | Alector Llc | Transferrin receptor antigen-binding domains and uses therefor |
WO2024023283A1 (en) | 2022-07-29 | 2024-02-01 | Institut National de la Santé et de la Recherche Médicale | Lrrc33 as a biomarker and biotarget in cutaneous t-cell lymphomas |
WO2024026447A1 (en) | 2022-07-29 | 2024-02-01 | Alector Llc | Anti-gpnmb antibodies and methods of use thereof |
WO2024026471A1 (en) | 2022-07-29 | 2024-02-01 | Alector Llc | Cd98hc antigen-binding domains and uses therefor |
WO2024028433A1 (en) | 2022-08-04 | 2024-02-08 | Institut National de la Santé et de la Recherche Médicale | Methods for the treatment of lymphoproliferative disorders |
WO2024033362A1 (en) | 2022-08-08 | 2024-02-15 | Atb Therapeutics | Humanized antibodies against cd79b |
WO2024033400A1 (en) | 2022-08-10 | 2024-02-15 | Institut National de la Santé et de la Recherche Médicale | Sk2 inhibitor for the treatment of pancreatic cancer |
WO2024033399A1 (en) | 2022-08-10 | 2024-02-15 | Institut National de la Santé et de la Recherche Médicale | Sigmar1 ligand for the treatment of pancreatic cancer |
WO2024040020A1 (en) | 2022-08-15 | 2024-02-22 | Absci Corporation | Quantitative affinity activity specific cell enrichment |
WO2024038112A1 (en) | 2022-08-17 | 2024-02-22 | Institut National de la Santé et de la Recherche Médicale | Improved anti-albumin nanobodies and their uses |
WO2024047110A1 (en) | 2022-08-31 | 2024-03-07 | Institut National de la Santé et de la Recherche Médicale | Method to generate more efficient car-t cells |
WO2024049949A1 (en) | 2022-09-01 | 2024-03-07 | Genentech, Inc. | Therapeutic and diagnostic methods for bladder cancer |
WO2024052503A1 (en) | 2022-09-08 | 2024-03-14 | Institut National de la Santé et de la Recherche Médicale | Antibodies having specificity to ltbp2 and uses thereof |
WO2024056668A1 (en) | 2022-09-12 | 2024-03-21 | Institut National de la Santé et de la Recherche Médicale | New anti-itgb8 antibodies and its uses thereof |
WO2024068705A1 (en) | 2022-09-29 | 2024-04-04 | F. Hoffmann-La Roche Ag | Protease-activated polypeptides |
WO2024074498A1 (en) | 2022-10-04 | 2024-04-11 | Imcheck Therapeutics | Combination of a btn3a activating antibody, a bcl2 inhibitor and hypomethylating agent for use in treating cancer |
WO2024077191A1 (en) | 2022-10-05 | 2024-04-11 | Flagship Pioneering Innovations V, Inc. | Nucleic acid molecules encoding trif and additionalpolypeptides and their use in treating cancer |
WO2024077239A1 (en) | 2022-10-07 | 2024-04-11 | Genentech, Inc. | Methods of treating cancer with anti-c-c motif chemokine receptor 8 (ccr8) antibodies |
WO2024074713A1 (en) | 2022-10-07 | 2024-04-11 | Institut National de la Santé et de la Recherche Médicale | Method to generate improving car-t cells |
Citations (34)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4356270A (en) * | 1977-11-08 | 1982-10-26 | Genentech, Inc. | Recombinant DNA cloning vehicle |
US4642334A (en) * | 1982-03-15 | 1987-02-10 | Dnax Research Institute Of Molecular And Cellular Biology, Inc. | Hybrid DNA prepared binding composition |
US4656134A (en) * | 1982-01-11 | 1987-04-07 | Board Of Trustees Of Leland Stanford Jr. University | Gene amplification in eukaryotic cells |
US4704692A (en) * | 1986-09-02 | 1987-11-03 | Ladner Robert C | Computer based system and method for determining and displaying possible chemical structures for converting double- or multiple-chain polypeptides to single-chain polypeptides |
US4711845A (en) * | 1984-08-31 | 1987-12-08 | Cetus Corporation | Portable temperature-sensitive control cassette |
US4714681A (en) * | 1981-07-01 | 1987-12-22 | The Board Of Reagents, The University Of Texas System Cancer Center | Quadroma cells and trioma cells and methods for the production of same |
US4800159A (en) * | 1986-02-07 | 1989-01-24 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences |
US4806471A (en) * | 1982-09-16 | 1989-02-21 | A/S Alfred Benzon | Plasmids with conditional uncontrolled replication behavior |
US4816397A (en) * | 1983-03-25 | 1989-03-28 | Celltech, Limited | Multichain polypeptides or proteins and processes for their production |
US4889818A (en) * | 1986-08-22 | 1989-12-26 | Cetus Corporation | Purified thermostable enzyme |
US4937193A (en) * | 1986-06-27 | 1990-06-26 | Delta Biotechnology Limited | Process for the genetic modification of yeast |
US4946786A (en) * | 1987-01-14 | 1990-08-07 | President And Fellows Of Harvard College | T7 DNA polymerase |
US4959317A (en) * | 1985-10-07 | 1990-09-25 | E. I. Du Pont De Nemours And Company | Site-specific recombination of DNA in eukaryotic cells |
US4965188A (en) * | 1986-08-22 | 1990-10-23 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme |
US4978743A (en) * | 1987-09-24 | 1990-12-18 | Bayer Aktiengesellschaft | Process for the continuous, counter flow/direct flow extraction of polyamides |
US4978745A (en) * | 1987-11-23 | 1990-12-18 | Centocor, Inc. | Immunoreactive heterochain antibodies |
US4983728A (en) * | 1987-07-31 | 1991-01-08 | Ire-Celltarg S.A. | Nucleic acid probes of human papilloma virus |
US5023171A (en) * | 1989-08-10 | 1991-06-11 | Mayo Foundation For Medical Education And Research | Method for gene splicing by overlap extension using the polymerase chain reaction |
US5030565A (en) * | 1983-08-17 | 1991-07-09 | Scripps Clinic And Research Foundation | Polypeptide-induced monoclonal receptors to protein ligands |
US5091513A (en) * | 1987-05-21 | 1992-02-25 | Creative Biomolecules, Inc. | Biosynthetic antibody binding sites |
US5126258A (en) * | 1984-09-07 | 1992-06-30 | Scripps Clinic And Research Foundation | Molecules with antibody combining sites that exhibit catalytic properties |
US5132405A (en) * | 1987-05-21 | 1992-07-21 | Creative Biomolecules, Inc. | Biosynthetic antibody binding sites |
US5225539A (en) * | 1986-03-27 | 1993-07-06 | Medical Research Council | Recombinant altered antibodies and methods of making altered antibodies |
US5229072A (en) * | 1992-02-03 | 1993-07-20 | Liquid Carbonic Inc. | Use of interhalogen compounds as a sterilizing agent |
US5229292A (en) * | 1986-07-28 | 1993-07-20 | Stine Seed Farm, Inc. | Biological control of insects using pseudomonas strains transformed with bacillus thuringiensis insect toxingene |
US5534254A (en) * | 1992-02-06 | 1996-07-09 | Chiron Corporation | Biosynthetic binding proteins for immuno-targeting |
US5846818A (en) * | 1985-11-01 | 1998-12-08 | Xoma Corporation | Pectate lyase signal sequence |
US5885793A (en) * | 1991-12-02 | 1999-03-23 | Medical Research Council | Production of anti-self antibodies from antibody segment repertoires and displayed on phage |
US5969108A (en) * | 1990-07-10 | 1999-10-19 | Medical Research Council | Methods for producing members of specific binding pairs |
US6207804B1 (en) * | 1987-05-21 | 2001-03-27 | Curis, Inc. | Genetically engineered antibody analogues and fusion proteins thereof |
US6214553B1 (en) * | 1997-01-21 | 2001-04-10 | Massachusetts General Hospital | Libraries of protein encoding RNA-protein fusions |
US6248516B1 (en) * | 1988-11-11 | 2001-06-19 | Medical Research Council | Single domain ligands, receptors comprising said ligands methods for their production, and use of said ligands and receptors |
US6291159B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for producing polymers having a preselected activity |
US6291158B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for tapping the immunological repertoire |
Family Cites Families (20)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4816567A (en) | 1983-04-08 | 1989-03-28 | Genentech, Inc. | Recombinant immunoglobin preparations |
JPS6147500A (en) | 1984-08-15 | 1986-03-07 | Res Dev Corp Of Japan | Chimera monoclonal antibody and its preparation |
EP0173494A3 (en) | 1984-08-27 | 1987-11-25 | The Board Of Trustees Of The Leland Stanford Junior University | Chimeric receptors by dna splicing and expression |
GB8422238D0 (en) | 1984-09-03 | 1984-10-10 | Neuberger M S | Chimeric proteins |
JPS61104788A (en) | 1984-10-26 | 1986-05-23 | Teijin Ltd | Nucleic acid base sequence |
DK171161B1 (en) | 1985-03-28 | 1996-07-08 | Hoffmann La Roche | A method for detecting the presence or absence of at least one specific nucleic acid sequence in a sample or for distinguishing two different nucleic acid sequences in this sample |
US4683195A (en) | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
DE3689123T2 (en) * | 1985-11-01 | 1994-03-03 | Xoma Corp | MODULAR UNIT OF ANTIBODY GENES, ANTIBODIES MADE THEREOF AND USE. |
GB8607679D0 (en) | 1986-03-27 | 1986-04-30 | Winter G P | Recombinant dna product |
SE458555B (en) | 1986-07-21 | 1989-04-10 | Atte Heikkilae | LINFAESTANORDNING |
JPS63152984A (en) | 1986-08-18 | 1988-06-25 | Wakunaga Pharmaceut Co Ltd | Dna coding l-chain of antipyocyanic human-type antibody |
DE3785186T2 (en) * | 1986-09-02 | 1993-07-15 | Enzon Lab Inc | BINDING MOLECULE WITH SINGLE POLYPEPTIDE CHAIN. |
DE3852304T3 (en) * | 1987-03-02 | 1999-07-01 | Enzon Lab Inc | Organism as carrier for "Single Chain Antibody Domain (SCAD)". |
CA1341235C (en) | 1987-07-24 | 2001-05-22 | Randy R. Robinson | Modular assembly of antibody genes, antibodies prepared thereby and use |
CA2016842A1 (en) | 1989-05-16 | 1990-11-16 | Richard A. Lerner | Method for tapping the immunological repertoire |
CA2057923A1 (en) | 1989-05-16 | 1990-11-17 | William D. Huse | Co-expression of heteromeric receptors |
CA2016841C (en) | 1989-05-16 | 1999-09-21 | William D. Huse | A method for producing polymers having a preselected activity |
AU627591B2 (en) | 1989-06-19 | 1992-08-27 | Xoma Corporation | Chimeric mouse-human km10 antibody with specificity to a human tumor cell antigen |
ES2176484T3 (en) | 1995-08-18 | 2002-12-01 | Morphosys Ag | PROTEIN BANKS / (POLI) PEPTIDES. |
-
1989
- 1989-11-13 EP EP89311731A patent/EP0368684B2/en not_active Expired - Lifetime
- 1989-11-13 AU AU45201/89A patent/AU634186B2/en not_active Expired
- 1989-11-13 DE DE68913658T patent/DE68913658T3/en not_active Expired - Lifetime
- 1989-11-13 AT AT89311731T patent/ATE102631T1/en not_active IP Right Cessation
- 1989-11-13 ES ES89311731T patent/ES2052027T5/en not_active Expired - Lifetime
- 1989-11-13 JP JP1511700A patent/JP2919890B2/en not_active Expired - Lifetime
- 1989-11-13 KR KR1019900701475A patent/KR0184860B1/en not_active IP Right Cessation
- 1989-11-13 WO PCT/GB1989/001344 patent/WO1990005144A1/en active Application Filing
- 1989-11-14 CA CA002002868A patent/CA2002868C/en not_active Expired - Lifetime
-
1990
- 1990-07-09 NO NO90903059A patent/NO903059L/en unknown
- 1990-07-09 DK DK199001647A patent/DK175392B1/en not_active IP Right Cessation
- 1990-07-10 FI FI903489A patent/FI903489A0/en not_active Application Discontinuation
-
1995
- 1995-06-06 US US08/470,031 patent/US6248516B1/en not_active Expired - Lifetime
-
2000
- 2000-11-28 US US09/722,364 patent/US6545142B1/en not_active Expired - Fee Related
-
2002
- 2002-11-08 US US10/290,252 patent/US7306907B2/en not_active Expired - Fee Related
- 2002-11-08 US US10/290,233 patent/US20040110941A2/en not_active Abandoned
-
2008
- 2008-05-27 US US12/127,237 patent/US20080299618A1/en not_active Abandoned
Patent Citations (36)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4356270A (en) * | 1977-11-08 | 1982-10-26 | Genentech, Inc. | Recombinant DNA cloning vehicle |
US4714681A (en) * | 1981-07-01 | 1987-12-22 | The Board Of Reagents, The University Of Texas System Cancer Center | Quadroma cells and trioma cells and methods for the production of same |
US4656134A (en) * | 1982-01-11 | 1987-04-07 | Board Of Trustees Of Leland Stanford Jr. University | Gene amplification in eukaryotic cells |
US4642334A (en) * | 1982-03-15 | 1987-02-10 | Dnax Research Institute Of Molecular And Cellular Biology, Inc. | Hybrid DNA prepared binding composition |
US4806471A (en) * | 1982-09-16 | 1989-02-21 | A/S Alfred Benzon | Plasmids with conditional uncontrolled replication behavior |
US4816397A (en) * | 1983-03-25 | 1989-03-28 | Celltech, Limited | Multichain polypeptides or proteins and processes for their production |
US5030565A (en) * | 1983-08-17 | 1991-07-09 | Scripps Clinic And Research Foundation | Polypeptide-induced monoclonal receptors to protein ligands |
US4711845A (en) * | 1984-08-31 | 1987-12-08 | Cetus Corporation | Portable temperature-sensitive control cassette |
US5126258A (en) * | 1984-09-07 | 1992-06-30 | Scripps Clinic And Research Foundation | Molecules with antibody combining sites that exhibit catalytic properties |
US4959317A (en) * | 1985-10-07 | 1990-09-25 | E. I. Du Pont De Nemours And Company | Site-specific recombination of DNA in eukaryotic cells |
US5846818A (en) * | 1985-11-01 | 1998-12-08 | Xoma Corporation | Pectate lyase signal sequence |
US4800159A (en) * | 1986-02-07 | 1989-01-24 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences |
US5225539A (en) * | 1986-03-27 | 1993-07-06 | Medical Research Council | Recombinant altered antibodies and methods of making altered antibodies |
US4937193A (en) * | 1986-06-27 | 1990-06-26 | Delta Biotechnology Limited | Process for the genetic modification of yeast |
US5229292A (en) * | 1986-07-28 | 1993-07-20 | Stine Seed Farm, Inc. | Biological control of insects using pseudomonas strains transformed with bacillus thuringiensis insect toxingene |
US4889818A (en) * | 1986-08-22 | 1989-12-26 | Cetus Corporation | Purified thermostable enzyme |
US4965188A (en) * | 1986-08-22 | 1990-10-23 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme |
US4704692A (en) * | 1986-09-02 | 1987-11-03 | Ladner Robert C | Computer based system and method for determining and displaying possible chemical structures for converting double- or multiple-chain polypeptides to single-chain polypeptides |
US4946786A (en) * | 1987-01-14 | 1990-08-07 | President And Fellows Of Harvard College | T7 DNA polymerase |
US5132405A (en) * | 1987-05-21 | 1992-07-21 | Creative Biomolecules, Inc. | Biosynthetic antibody binding sites |
US6207804B1 (en) * | 1987-05-21 | 2001-03-27 | Curis, Inc. | Genetically engineered antibody analogues and fusion proteins thereof |
US5091513A (en) * | 1987-05-21 | 1992-02-25 | Creative Biomolecules, Inc. | Biosynthetic antibody binding sites |
US4983728A (en) * | 1987-07-31 | 1991-01-08 | Ire-Celltarg S.A. | Nucleic acid probes of human papilloma virus |
US4978743A (en) * | 1987-09-24 | 1990-12-18 | Bayer Aktiengesellschaft | Process for the continuous, counter flow/direct flow extraction of polyamides |
US4978745A (en) * | 1987-11-23 | 1990-12-18 | Centocor, Inc. | Immunoreactive heterochain antibodies |
US20030130496A1 (en) * | 1988-11-11 | 2003-07-10 | Medical Research Council | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US6545142B1 (en) * | 1988-11-11 | 2003-04-08 | Medical Research Council Of The United Kingdom | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US6248516B1 (en) * | 1988-11-11 | 2001-06-19 | Medical Research Council | Single domain ligands, receptors comprising said ligands methods for their production, and use of said ligands and receptors |
US6291159B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for producing polymers having a preselected activity |
US6291158B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for tapping the immunological repertoire |
US5023171A (en) * | 1989-08-10 | 1991-06-11 | Mayo Foundation For Medical Education And Research | Method for gene splicing by overlap extension using the polymerase chain reaction |
US5969108A (en) * | 1990-07-10 | 1999-10-19 | Medical Research Council | Methods for producing members of specific binding pairs |
US5885793A (en) * | 1991-12-02 | 1999-03-23 | Medical Research Council | Production of anti-self antibodies from antibody segment repertoires and displayed on phage |
US5229072A (en) * | 1992-02-03 | 1993-07-20 | Liquid Carbonic Inc. | Use of interhalogen compounds as a sterilizing agent |
US5534254A (en) * | 1992-02-06 | 1996-07-09 | Chiron Corporation | Biosynthetic binding proteins for immuno-targeting |
US6214553B1 (en) * | 1997-01-21 | 2001-04-10 | Massachusetts General Hospital | Libraries of protein encoding RNA-protein fusions |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8940871B2 (en) | 2006-03-20 | 2015-01-27 | The Regents Of The University Of California | Engineered anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting |
US8940298B2 (en) | 2007-09-04 | 2015-01-27 | The Regents Of The University Of California | High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection |
US9527919B2 (en) | 2007-09-04 | 2016-12-27 | The Regents Of The University Of California | High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection |
Also Published As
Publication number | Publication date |
---|---|
ES2052027T3 (en) | 1994-07-01 |
NO903059L (en) | 1990-09-07 |
DE68913658D1 (en) | 1994-04-14 |
KR920700228A (en) | 1992-02-19 |
WO1990005144A1 (en) | 1990-05-17 |
DK164790D0 (en) | 1990-07-09 |
DE68913658T3 (en) | 2005-07-21 |
US6545142B1 (en) | 2003-04-08 |
EP0368684A1 (en) | 1990-05-16 |
DK164790A (en) | 1990-09-07 |
JP2919890B2 (en) | 1999-07-19 |
US7306907B2 (en) | 2007-12-11 |
AU4520189A (en) | 1990-05-28 |
ATE102631T1 (en) | 1994-03-15 |
US6248516B1 (en) | 2001-06-19 |
EP0368684B1 (en) | 1994-03-09 |
DE68913658T2 (en) | 1994-09-08 |
NO903059D0 (en) | 1990-07-09 |
US20030130496A1 (en) | 2003-07-10 |
FI903489A0 (en) | 1990-07-10 |
DK175392B1 (en) | 2004-09-20 |
US20040110941A2 (en) | 2004-06-10 |
AU634186B2 (en) | 1993-02-18 |
KR0184860B1 (en) | 1999-04-01 |
CA2002868C (en) | 2007-03-20 |
CA2002868A1 (en) | 1990-05-11 |
ES2052027T5 (en) | 2005-04-16 |
US20030114659A1 (en) | 2003-06-19 |
JPH03502801A (en) | 1991-06-27 |
EP0368684B2 (en) | 2004-09-29 |
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