Transgenic mammals having human Ig loci including plural VHand VKregions and antibodies produced therefrom

• US 7,064,244 B2
• Filed: 02/19/2002
• Issued: 06/20/2006
• Est. Priority Date: 12/03/1996
• Status: Expired due to Fees

• Current Assignee: Abgenix Inc (Amgen Inc.)
• Original Assignee: Abgenix Inc (Amgen Inc.)
• Inventors: Jakobovits, Aya, Green, Larry, Kucherlapati, Raju, Klapholz, Susan, Mendez, Michael J
• Primary Examiner(s): Wehbe′, Anne M.
• Application Number:
  US10/078,958
• Publication Number:
  US 20030070185A1
• Time in Patent Office: 1,582 Days
• Field of Search: 800/13, 800/18
• US Class Current: 800/18
• 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 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C07K 14/415 from plants
• 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 16/005 constructed by phage libraries
• CPC Class Codes: C07K 16/241 Tumor Necrosis Factors
• CPC Class Codes: C07K 16/244 Interleukins [IL]
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• 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 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12N 15/8509 : for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2800/206 : of yeast origin, e.g. YAC, 2u

Abstract

The present invention relates to transgenic non-human animals that are engineered to contain human immunoglobulin gene loci. In particular, animals in accordance with the invention possess human Ig loci that include plural variable (V_H and Vκ) gene regions. Advantageously, the inclusion of plural variable region genes enhances the specificity and diversity of human antibodies produced by the animal. Further, the inclusion of such regions enhances and reconstitutes B-cell development to the animals, such that the animals possess abundant mature B-cells secreting extremely high affinity antibodies.

Claims

1. A transgenic mouse whose genome comprises a fragment of human chromosome 14 from the five most proximal V_H gene segments, continuing through the D segment genes, the J segment genes and the constant region genes through Cδ

of the human immunoglobulin heavy chain locus, wherein said fragment does not contain a Cγ

gene, and wherein said fragment is operably linked to a human Cγ

gene selected from the group consisting of;

a human Cγ

1 gene, a human Cγ

3 gene, and a human Cγ

4 gene

said transgenic mouse producing fully human antibodies having IgG1, IgG3, or IgG4 heavy chains, respectively, specific for a desired antigen when immunized with said desired antigen.

2. The transgenic mouse according to claim 1, wherein said mouse further comprises a fragment of human chromosome 2 comprising human V_κ

, human J_κ

and human C_κ

gene segments of an immunoglobulin kappa light chain locus

said transgenic mouse producing fully human antibodies having IgG1, IgG3, or IgG4 heavy chains, respectively, specific for a desired antigen when immunized with said desired antigen.

3. The transgenic mouse according to claim 2, wherein said fragment of human chromosome 2 extends from the three most proximal V_κ

gene segments, continuing through the J_κ

and C_κ

gene segments, through the human kappa deleting element.

4. The transgenic mouse according to any one of claims 1–

3, wherein said mouse further comprises;

a) inactivated endogenous immunoglobulin heavy chain loci in which all of the J segment genes are deleted to prevent rearrangement and to prevent formation of a transcript of a rearranged locus and the expression of an endogenous immunoglobulin heavy chain; and

b) inactivated endogenous immunoglobulin light chain loci in which the C_κ

gene is deleted to prevent rearrangement and to prevent formation of a transcript of a rearranged locus and the expression of an endogenous immunoglobulin light chain;

wherein said transgenic mouse lacks expression of endogenous immunoglobulin heavy and kappa light chains.

5. A method for producing a fully human IgG antibody specific for a desired antigen, comprising:

(a) immunizing a transgenic mouse according to any one of claims 1–

3 with said desired antigen; and

(b) recovering the antibody.

6. A method for producing a fully human IgG antibody specific for a desired antigen, comprising:

(a) immunizing a transgenic mouse according to claim 4 with said desired antigen; and

(b) recovering the antibody.