Human antibodies derived from immunized xenomice

• US 6,657,103 B1
• Filed: 09/04/1997
• Issued: 12/02/2003
• Est. Priority Date: 01/12/1990
• Status: Expired due to Fees

• Current Assignee: Abgenix Inc (Amgen Inc.)
• Original Assignee: Abgenix Inc (Amgen Inc.)
• Inventors: Brenner, Daniel G., Jakobovits, Aya, Capon, Daniel J., Kucherlapati, Raju, Klapholz, Sue
• Primary Examiner(s): Wehbe′ , Anne M.
• Application Number:
  US08/923,138
• Time in Patent Office: 2,280 Days
• Field of Search: 800/4, 800/6, 800/18, 435/320.1
• US Class Current: 800/6
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/072 maintaining or altering function, i.e. knock in
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 2267/0381 Animal model for diseases of the hematopoietic system
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/1282 from Clostridium (G)
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/241 Tumor Necrosis Factors
• CPC Class Codes: C07K 16/244 Interleukins [IL]
• CPC Class Codes: C07K 16/248 IL-6
• CPC Class Codes: C07K 16/2812 against CD4
• CPC Class Codes: C07K 16/2854 against selectins, e.g. CD62
• CPC Class Codes: C07K 16/2875 against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 against NGF C07K16/22, against TNF C07K16/241
• CPC Class Codes: C07K 16/462 Igs containing a variable region (Fv) from one specie and a constant region (Fc) from another
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]
• CPC Class Codes: C12N 15/8509 : for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 15/87 : Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 15/90 : Stable introduction of foreign DNA into chromosome
• CPC Class Codes: C12N 2840/203 : having an IRES

Abstract

Antibodies with fully human variable regions against a specific antigen can be prepared by administering the antigen to a transgenic animal which has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled. Various subsequent manipulations can be performed to obtain either antibodies per se or analogs thereof.

Claims

1. A method for producing a human IgM comprising a nonchimeric variable region, wherein said immunoglobulin is specific for a desired antigen, wherein said method comprises the steps of:

(a) administering said antigen or an immunogenic portion thereof to a transgenic mouse under conditions to stimulate an immune response, whereby said animal produces B cells that produce said immunoglobulin specific for said antigen,

wherein the genome of said transgenic mouse is characterized by inactivated endogenous immunoglobulin heavy chain loci in which all of the J segment genes are deleted to prevent rearrangement of the loci and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, and by inactivated endogenous immunoglobulin light chain loci in which genes are deleted to prevent rearrangement of the loci and to prevent formation of a transcript of a rearranged immunoglobulin light chain locus,

and wherein said genome of said transgenic mouse further comprises a DNA fragment of a human immunoglobulin heavy chain locus, the fragment being a SpeI-SpeI fragment commencing from the VH6 gene and continuing through the human D segment genes, human J segment genes and human constant region genes and into the Cδ

gene of that locus, wherein said SpeI-SpeI fragment does not include a gamma constant region, and further comprises a fragment of human chromosome 2, said fragment comprising Vκ

, Jκ and

Cκ

gene segments of a human immunoglobulin kappa light chain locus, and

(b) recovering said immunoglobulin, provided that if said immunoglobulin comprises a human constant region, said human constant region is a human mu constant region or a human kappa constant region, or both.

3. The method according to claim 1 or 2, wherein said recovering step comprises recovering polyclonal immunoglobulin from said transgenic mouse.

4. The method according to claim 1 or 2, wherein said recovering step comprises immortalizing B cells from said transgenic mouse immunized with said antigen, screening the resulting immortalized cells for the secretion of said immunoglobulin specific for said antigen, and

(a) recovering immunoglobulin secreted by said immortalized B cells, or

(b) recovering the genes encoding at least the variable region of said immunoglobulin from the immortalized B cells, and optionally modifying said genes to encode a modified immunoglobulin;

(c) expressing said genes or modified forms thereof to produce said immunoglobulin or said modified immunoglobulin; and

(d) recovering said immunoglobulin or said modified immunoglobulin.

2. A method for producing an antigen binding fragment of a human IgM, said fragment comprising a nonchimeric variable region, wherein said fragment is specific for a desired antigen, wherein said method comprises the steps of:

(a) administering said antigen or an immunogenic portion thereof to a transgenic mouse under conditions to stimulate an immune response, whereby said animal produces B cells that produce said immunoglobulin specific for said antigen,

wherein the genome of said transgenic mouse is characterized by inactivated endogenous immunoglobulin heavy chain loci in which all of the J segment genes are deleted to prevent rearrangement of the loci and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, by inactivated endogenous immunoglobulin light chain loci in which one or more genes are deleted to prevent rearrangement of the loci and to prevent formation of a transcript of a rearranged immunoglobulin light chain locus,

and wherein said genome of said transgenic mouse further comprises a DNA fragment of a human immunoglobulin heavy chain locus, the fragment being a SpeI-SpeI fragment commencing from the VH6 gene and continuing through the human D segment genes, human J segment genes and human constant region genes and into the Cδ

gene of that locus, wherein said SpeI-SpeI fragment does not include a gamma constant region, and further comprises a fragment of human chromosome 2, said fragment comprising Vκ

, Jκ and

Cκ

gene segments of a human immunoglobulin kappa light chain locus,

(b) recovering said immunoglobulin specific for said antigen, and

(c) producing an antigen-binding fragment from said immunoglobulin recovered in step (b).