A method of screening is provided. In certain embodiments, the method involves a) obtaining the nucleotide sequences of: i. a heavy chain-encoding nucleic acid that encodes the variable domain of a heavy chain of a first antibody of an animal; and ii. a light chain-encoding nucleic acid that encodes the variable domain of a light chain of the first antibody; b) obtaining nucleotide sequences of cDNAs encoding at least a portion of the antibody repertoire of the animal; c) computationally screening the sequences obtained in b) to identify heavy and light chain sequences that are related by lineage to the heavy and light chain sequences of a); and d) testing at least one pair of the heavy and light chain sequences identified in c) to identify a second antibody that binds to the same antigen as the first antibody.
C07K 16/32 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products from oncogenes
Provided herein, inter alia, is a method for producing an enriched population of antigen-specific plasma cells. In some embodiments, the method may comprise: (a) obtaining a sample of cells from an animal that has been immunized by an antigen, wherein the sample comprises B cells; (b) enriching for a population of antigen-specific B cells that comprise cell surface antibodies that are specific for the antigen by: i. contacting at least 105 of the cells in said sample, en masse, with the antigen or a portion thereof; and ii. isolating cells that bind to the antigen or portion thereof; and (c) activating the enriched B cells, en masse, in the presence of the antigen or portion thereof, to produce the enriched population of antigen-specific plasma cells.
Provided herein, inter alia, is a method for producing an enriched population of antigen-specific plasma cells. In some embodiments, the method may comprise: (a) obtaining a sample of cells from an animal that has been immunized by an antigen, wherein the sample comprises B cells; (b) enriching for a population of antigen-specific B cells that comprise cell surface antibodies that are specific for the antigen by: i. contacting at least 105 of the cells in said sample, en masse, with the antigen or a portion thereof; and ii. isolating cells that bind to the antigen or portion thereof; and (c) activating the enriched B cells, en masse, in the presence of the antigen or portion thereof, to produce the enriched population of antigen-specific plasma cells.
The invention provides a rabbit-derived immortal B-lymphocyte capable of fusion with a rabbit splenocyte to produce a hybrid cell that produces an antibody. The immortal B-lymphocyte does not detectably express endogenous immunoglobulin heavy chain and may contain, in certain embodiments, an altered immunoglobulin heavy chain-encoding gene. A hybridoma resulting from fusion between the subject immortal B-lymphocyte and a rabbit antibody-producing cell is provided, as is a method of using that hybridoma to produce an antibody. The subject invention finds use in a variety of different diagnostic, therapeutic and research applications.
In certain embodiments, the method may comprise: a) obtaining the antibody sequences from a population of B cells; b) grouping the antibody sequences to provide a plurality of groups of lineage-related antibodies; c) testing a single antibody from each of the groups in a bioassay and, after the first antibody has been identified, d) testing further antibodies that are in the same group as the first antibody in a second bioassay. In another embodiment, the method may comprise: a) testing a plurality of antibodies obtained from a first portion of an antibody producing organ of an animal; b) obtaining the sequence of a first identified antibody; c) obtaining from a second portion of said antibody producing organ the sequences of further antibodies that are related by lineage to said first antibody; and, c) testing the further antibodies in a second bioassay.
C07K 16/28 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
G01N 33/68 - Chemical analysis of biological material, e.g. blood, urineTesting involving biospecific ligand binding methodsImmunological testing involving proteins, peptides or amino acids
G01N 33/566 - ImmunoassayBiospecific binding assayMaterials therefor using specific carrier or receptor proteins as ligand binding reagent
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
A method of screening is provided. In certain embodiments, the method involves a) obtaining the nucleotide sequences of: i. a heavy chain-encoding nucleic acid that encodes the variable domain of a heavy chain of a first antibody of an animal; and ii. a light chain-encoding nucleic acid that encodes the variable domain of a light chain of the first antibody; b) obtaining nucleotide sequences of cDNAs encoding at least a portion of the antibody repertoire of the animal; c) computationally screening the sequences obtained in b) to identify heavy and light chain sequences that are related by lineage to the heavy and light chain sequences of a); and d) testing at least one pair of the heavy and light chain sequences identified in c) to identify a second antibody that binds to the same antigen as the first antibody.
The invention provides a method for identifying positions of an antibody that can be modified without significantly reducing the binding activity of the antibody. In many embodiments, the method involves identifying a substitutable position in a parent antibody by comparing its amino acid sequence to the amino acid sequences of a number of related antibodies that each bind to the same antigen as the parent antibody. The amino acid at the substitutable position may be substituted for a different amino acid without significantly affecting the activity of the antibody. The subject methods may be employed to change the amino acid sequence of a CDR without significantly reducing the affinity of the antibody of the antibody, in humanization methods, or in other antibody engineering methods. The invention finds use in a variety of therapeutic, diagnostic and research applications.
C07K 16/18 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies against material from animals or humans
C07K 16/24 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
In certain embodiments, the method may comprise: a) obtaining the antibody sequences from a population of B cells; b) grouping the antibody sequences to provide a plurality of groups of lineage-related antibodies; c) testing a single antibody from each of the groups in a bioassay and, after the first antibody has been identified, d) testing further antibodies that are in the same group as the first antibody in a second bioassay. In another embodiment, the method may comprise: a) testing a plurality of antibodies obtained from a first portion of an antibody producing organ of an animal; b) obtaining the sequence of a first identified antibody; c) obtaining from a second portion of said antibody producing organ the sequences of further antibodies that are related by lineage to said first antibody; and, c) testing the further antibodies in a second bioassay.
C07K 16/28 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
G01N 33/68 - Chemical analysis of biological material, e.g. blood, urineTesting involving biospecific ligand binding methodsImmunological testing involving proteins, peptides or amino acids
G01N 33/566 - ImmunoassayBiospecific binding assayMaterials therefor using specific carrier or receptor proteins as ligand binding reagent
9.
Fusion partner for production of monoclonal rabbit antibodies
The invention provides a rabbit-derived immortal B-lymphocyte capable of fusion with a rabbit splenocyte to produce a hybrid cell that produces an antibody. The immortal B-lymphocyte does not detectably express endogenous immunoglobulin heavy chain and may contain, in certain embodiments, an altered immunoglobulin heavy chain-encoding gene. A hybridoma resulting from fusion between the subject immortal B-lymphocyte and a rabbit antibody-producing cell is provided, as is a method of using that hybridoma to produce an antibody. The subject invention finds use in a variety of different diagnostic, therapeutic and research applications.
In certain embodiments, the method may comprise: a) obtaining the antibody sequences from a population of B cells; b) grouping the antibody sequences to provide a plurality of groups of lineage-related antibodies; c) testing a single antibody from each of the groups in a bioassay and, after the first antibody has been identified, d) testing further antibodies that are in the same group as the first antibody in a second bioassay. In another embodiment, the method may comprise: a) testing a plurality of antibodies obtained from a first portion of an antibody producing organ of an animal; b) obtaining the sequence of a first identified antibody; c) obtaining from a second portion of said antibody producing organ the sequences of further antibodies that are related by lineage to said first antibody; and, c) testing the further antibodies in a second bioassay.
C07K 16/28 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
G01N 33/68 - Chemical analysis of biological material, e.g. blood, urineTesting involving biospecific ligand binding methodsImmunological testing involving proteins, peptides or amino acids
G01N 33/566 - ImmunoassayBiospecific binding assayMaterials therefor using specific carrier or receptor proteins as ligand binding reagent
11.
FACS-based method for obtaining an antibody sequence
In certain embodiments, the method may comprises: a) contacting a population of permeabilized, cross-linked antibody-producing cells with a labeled antigen to produce a labeled sample in which cells that produce an antibody that specifically binds to said antigen are intracellularly labeled; b) using FACS to isolate cells that are intracellularly labeled, thereby producing labeled cells; c) uncrosslinking said labeled cells to produce uncrosslinked cells; and d) amplifying heavy and light chain-encoding nucleic acid from individual uncrosslinked cells, thereby obtaining nucleic acid that encodes the variable domain of antibody that specifically binds to said antigen.
In certain embodiments, the method may comprises: a) contacting a population of permeabilized, cross-linked antibody-producing cells with a labeled antigen to produce a labeled sample in which cells that produce an antibody that specifically binds to said antigen are intracellularly labeled; b) using FACS to isolate cells that are intracellularly labeled, thereby producing labeled cells; c) uncrosslinking said labeled cells to produce uncrosslinked cells; and d) amplifying heavy and light chain-encoding nucleic acid from individual uncrosslinked cells, thereby obtaining nucleic acid that encodes the variable domain of antibody that specifically binds to said antigen.
01 - Chemical and biological materials for industrial, scientific and agricultural use
05 - Pharmaceutical, veterinary and sanitary products
Goods & Services
Biochemicals, namely, rabbit monoclonal antibodies for in
vitro scientific or research use. Diagnostic kits consisting primarily of rabbit monoclonal
antibodies, buffers, and reagents for use in disease
testing.
In certain embodiments, the method may comprise: a) obtaining the antibody sequences from a population of B cells; b) grouping the antibody sequences to provide a plurality of groups of lineage-related antibodies; c) testing a single antibody from each of the groups in a bioassay and, after the first antibody has been identified, d) testing further antibodies that are in the same group as the first antibody in a second bioassay. In another embodiment, the method may comprise: a) testing a plurality of antibodies obtained from a first portion of an antibody producing organ of an animal; b) obtaining the sequence of a first identified antibody; c) obtaining from a second portion of said antibody producing organ the sequences of further antibodies that are related by lineage to said first antibody; and, c) testing the further antibodies in a second bioassay.
The invention provides a rabbit-derived immortal B-lymphocyte capable of fusion with a rabbit splenocyte to produce a hybrid cell that produces an antibody. The immortal B-lymphocyte does not detectably express endogenous immunoglobulin heavy chain and may contain, in certain embodiments, an altered immunoglobulin heavy chain-encoding gene. A hybridoma resulting from fusion between the subject immortal B-lymphocyte and a rabbit antibody-producing cell is provided, as is a method of using that hybridoma to produce an antibody. The subject invention finds use in a variety of different diagnostic, therapeutic and research applications.
In certain embodiments, the method may comprise: a) obtaining the antibody sequences from a population of B cells; b) grouping the antibody sequences to provide a plurality of groups of lineage-related antibodies; c) testing a single antibody from each of the groups in a bioassay and, after the first antibody has been identified, d) testing further antibodies that are in the same group as the first antibody in a second bioassay. In another embodiment, the method may comprise: a) testing a plurality of antibodies obtained from a first portion of an antibody producing organ of an animal; b) obtaining the sequence of a first identified antibody; c) obtaining from a second portion of said antibody producing organ the sequences of further antibodies that are related by lineage to said first antibody; and, c) testing the further antibodies in a second bioassay.
JIANGSU SIMCERE PHARMACEUTICAL R & D CO., LTD. (China)
EPITOMICS, INC (USA)
Inventor
Li, Pierre
Ke, Yaohuang
Zhang, Yongke
Zhu, Weimin
Yu, Guoliang
Ma, Fanxin
Fan, Xin
Abstract
The present invention provides an anti-VEGF monoclonal antibody, which has the variable region of heavy chain comprising the amino acid sequences of SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3, and/or the variable region of light chain comprising the amino acid sequences of SEQ ID NO.4, SEQ ID NO.5 and SEQ ID NO.6. The antibody can be produced by the cell line with the preservation number of CGMCC NO.3233. The invention also provides the use of said antibody for the manufacture of medicaments for the treatment of a disease that is relevant to VEGF, wherein further provided are pharmaceutical composition, agents, kits and chips comprising said antibody, as well as the cell line with the preservation number of CGMCC NO.3233.
The invention provides a rabbit-derived immortal B-lymphocyte capable of fusion with a rabbit splenocyte to produce a hybrid cell that produces an antibody. The immortal B-lymphocyte does not detectably express endogenous immunoglobulin heavy chain and may contain, in certain embodiments, an altered immunoglobulin heavy chain-encoding gene. A hybridoma resulting from fusion between the subject immortal B-lymphocyte and a rabbit antibody-producing cell is provided, as is a method of using that hybridoma to produce an antibody. The subject invention finds use in a variety of different diagnostic, therapeutic and research applications.
An antibody is provided. In certain cases, the antibody comprises: a) a heavy chain variable domain that comprises CDR regions that are substantially identical to the heavy chain CDR regions of a selected antibody and b) a light chain variable domain that comprises CDR regions that are substantially identical to the light chain CDR regions of the selected antibody, where the antibody binds a selected target.
A method of obtaining nucleic acid encoding a plurality of antibodies is provided. In certain embodiments, the method comprises obtaining from an immunized animal nucleic acid encoding the amino acid sequence of the heavy and light chains of a second antibody that binds to the antigen as a first antibody and differs in amino acid sequence to the first antibody, wherein the obtaining is done by amplification using: i. a first primer pair that includes oligonucleotides are complementary to CDR-encoding regions first antibody.
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
C40B 30/04 - Methods of screening libraries by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
C07K 16/00 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies
C07K 16/24 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
An antibody that binds TNF α and neutralizes its activity is provided. In certain cases, the antibody comprises: a) a heavy chain variable domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO. 1, and b) a light chain variable domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID No. 2.
A61K 39/395 - AntibodiesImmunoglobulinsImmune serum, e.g. antilymphocytic serum
C07K 16/24 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
An antibody that binds TNFα and neutralizes its activity is provided. In certain cases, the antibody comprises: a) a heavy chain variable domain comprising an amino acid sequence that is at least 95 % identical to the amino acid sequence of SEQ ID NO. 1, and b) a light chain variable domain comprising an amino acid sequence that is at least 95 % identical to the amino acid sequence of SEQ ID No. 2.
An antibody is provided. In certain cases, the antibody comprises: a) a heavy chain variable domain that comprises CDR regions that are substantially identical to the heavy chain CDR regions of a selected antibody and b) a light chain variable domain that comprises CDR regions that are substantially identical to the light chain CDR regions of the selected antibody, where the antibody binds a selected target.
An antibody is provided. In certain cases, the antibody comprises: a) a heavy chain variable domain that comprises CDR regions that are substantially identical to the heavy chain CDR regions of a selected antibody and b) a light chain variable domain that comprises CDR regions that are substantially identical to the light chain CDR regions of the selected antibody, where the antibody binds a selected target.
The invention provides monoclonal antibodies that neutralize TNFα activity. The monoclonal antibodies may be rabbit monoclonal antibodies or monoclonal antibodies having CDR regions derived from those rabbit monoclonal antibodies. In certain embodiments, the monoclonal antibodies may be humanized. Methods of using the subject antibodies to inhibit TNFα activity, methods of treatment using those antibodies and kits containing the same are also provided. The invention finds use in a variety of research and medical applications.
The invention provides a rabbit-derived immortal B-lymphocyte capable of fusion with a rabbit splenocyte to produce a hybrid cell that produces an antibody. The immortal B-lymphocyte does not detectably express endogenous immunoglobulin heavy chain and may contain, in certain embodiments, an altered immunoglobulin heavy chain-encoding gene. A hybridoma resulting from fusion between the subject immortal B-lymphocyte and a rabbit antibody-producing cell is provided, as is a method of using that hybridoma to produce an antibody. The subject invention finds use in a variety of different diagnostic, therapeutic and research applications.
The invention provides a rabbit-derived immortal B-lymphocyte capable of fusion with a rabbit splenocyte to produce a hybrid cell that produces an antibody. The immortal B-lymphocyte does not detectably express endogenous immunoglobulin heavy chain and may contain, in certain embodiments, an altered immunoglobulin heavy chain-encoding gene. A hybridoma resulting from fusion between the subject immortal B-lymphocyte and a rabbit antibody-producing cell is provided, as is a method of using that hybridoma to produce an antibody. The subject invention finds use in a variety of different diagnostic, therapeutic and research applications.
The invention provides monoclonal antibodies that neutralize TNFα activity. The monoclonal antibodies may be rabbit monoclonal antibodies or monoclonal antibodies having CDR regions derived from those rabbit monoclonal antibodies. In certain embodiments, the monoclonal antibodies may be humanized. Methods of using the subject antibodies to inhibit TNFα activity, methods of treatment using those antibodies and kits containing the same are also provided. The invention finds use in a variety of research and medical applications.
A61K 39/395 - AntibodiesImmunoglobulinsImmune serum, e.g. antilymphocytic serum
C07K 16/24 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
C12N 1/00 - Microorganisms, e.g. protozoaCompositions thereofProcesses of propagating, maintaining or preserving microorganisms or compositions thereofProcesses of preparing or isolating a composition containing a microorganismCulture media therefor
C12N 15/00 - Mutation or genetic engineeringDNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purificationUse of hosts therefor
01 - Chemical and biological materials for industrial, scientific and agricultural use
42 - Scientific, technological and industrial services, research and design
Goods & Services
(1) Biochemicals, namely, monoclonal antibodies for diagnostic and research use. (1) Technical consultation and research in the field of biochemistry, namely the development of technology for the production of monoclonal antibodies and the production of monoclonal antibodies for research, diagnostic, and therapeutic purposes.
01 - Chemical and biological materials for industrial, scientific and agricultural use
05 - Pharmaceutical, veterinary and sanitary products
42 - Scientific, technological and industrial services, research and design
Goods & Services
Chemicals used in industry, science and photography, as well as in agriculture, horticulture and forestry; unprocessed artificial resins, unprocessed plastics; manures; fire extinguishing compositions; tempering and soldering preparations; chemical substances for preserving foodstuffs; tanning substances; adhesives used in industry; biochemicals, including monoclonal antibodies for diagnostic and research use. Pharmaceutical and veterinary preparations; sanitary preparations for medical purposes; dietetic substances adapted for medical use, food for babies; plasters, materials for dressings; material for stopping teeth, dental wax; disinfectants; preparations for destroying vermin; fungicides, herbicides; biochemicals, including monoclonal antibodies for therapeutic use. Technical consultation and research in the field of biochemistry, namely the development of technology for the production of monoclonal antibodies and the production of monoclonal antibodies for research, diagnostic, and therapeutic purposes.
