Transgenic non-human animals capable of producing heterologous antibodies

• US 7,041,871 B1
• Filed: 11/28/2000
• Issued: 05/09/2006
• Est. Priority Date: 10/10/1995
• Status: Expired due to Term

• Current Assignee: Genpharm Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: Genpharm Incorporated (Bristol-Myers Squibb Company)
• Inventors: Kay, Robert M., Lonberg, Nils
• Primary Examiner(s): WEHBE, ANNE MARIE SABRINA
• Application Number:
  US09/724,965
• Time in Patent Office: 1,988 Days
• Field of Search: 800/13, 800/18, 435/320.1, 435/325, 435/455
• US Class Current: 800/18
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• 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 16/462 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 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic

Abstract

The invention relates to transgenic non-human animals capable of producing heterologous antibodies and methods for producing human sequence antibodies which bind to human antigens with substantial affinity.

Claims

1. A transgenic mouse comprising in its genome a human kappa light chain immunoglobulin transgene, said transgene containing five human light chain Vκ

segments, wherein the five human light chain Vκ

segments are L15, L18, L24, A10 and A27, a plurality of human light chain Jκ

segments, and a human light chain Cκ

segment, which segments are operably linked to transcription regulatory sequences and undergo rearrangement in B lymphocytes in vivo to produce a repertoire of rearranged transgenes encoding a plurality of human kappa light chain polypeptides, which human kappa light chain polypeptides are produced in said transgenic mouse.

2. The transgenic mouse of claim 1, wherein said transgene further comprises a human 3′

kappa enhancer segment.

3. The transgenic mouse of claim 2, wherein the human 3′

kappa enhancer segment is a 4 kb BamHI fragment containing the human 3′

kappa enhancer.

4. The transgenic mouse of claim 1, wherein said mouse further comprises an inactivated endogenous mouse kappa light chain immunoglobulin gene locus.

10. The transgenic mouse of claim 1, wherein the human kappa light chain immunoglobulin transgene is created by co-injection into a pronucleus of a mouse embryo of a yeast artificial chromosome (YAC) comprising multiple human Vκ

segments and at least one human immunoglobulin transgene comprising at least one human Vκ

segment, wherein the mouse embryo develops into said transgenic mouse.

11. The transgenic mouse of claim 10, wherein the YAC comprises at least 32 different Vκ

segments.

12. The transgenic mouse of claim 10, wherein the kappa light chain immunoglobulin transgene is created by co-injection into a pronucleus of a mouse embryo of a YAC comprising multiple human Vκ

segments, a first human immunoglobulin transgene comprising four human Vκ

segments and a second human immunoglobulin transgene comprising one human Vκ

segment, five human Jκ

segments, a human intronic enhancer, a human Cκ and

a human 3′

kappa enhancer.

13. The transgenic mouse of claim 1, wherein said human kappa light chain immunoglobulin transgene is KCo5.

5. A transgenic mouse comprising in its genome a human kappa light chain immunoglobulin transgene, said kappa light chain transgene containing five human light chain Vκ

segments, wherein the five human light chain Vκ

segments are L15, L18, L24, A10 and A27, a plurality of human light chain Jκ

segments, and a human light chain Cκ

segment, which segments are operably linked to transcription regulatory sequences and undergo rearrangement in B lymphocytes in vivo to produce a repertoire of rearranged transgenes encoding a plurality of human kappa light chain polypeptides, which human kappa light chain polypeptides are produced in said mouse, wherein said transgenic mouse further comprises a human heavy chain immunoglobulin transgene that produces a repertoire of human heavy chain polypeptides that pair with said kappa light chain polypeptides to form a repertoire of human immunoglobulins in said mouse.

6. The transgenic mouse of claim 5, wherein said human kappa light chain immunoglobulin transgene further comprises a human 3′

kappa enhancer segment.

7. The transgenic mouse of claim 6, wherein the human 3′

kappa enhancer segment is a 4 kb BamHI fragment containing the human 3′

kappa enhancer.

8. The transgenic mouse of claim 5, wherein said mouse further comprises an inactivated endogenous mouse kappa light chain immunoglobulin gene locus and an inactivated endogenous mouse heavy chain immunoglobulin gene locus.

9. The transgenic mouse of claim 6, which produces antigen-specific human immunoglobulins when said transgenic mouse is immunized with an antigen.

14. The transgenic mouse of claim 6, wherein said human kappa light chain immunoglobulin transgene is KCo5.