Human Lambda Light Chain Mice

• US 20120070861A1
• Filed: 06/22/2011
• Published: 03/22/2012
• Est. Priority Date: 06/22/2010
• Status: Active Grant

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: Regeneron Pharmaceuticals Incorporated
• Inventors: Macdonald, Lynn, Stevens, Sean, Gurer, Cagan, Murphy, Andrew J., Hosiawa, Karolina A.
• Granted Patent: US 9,012,717 B2
• Time in Patent Office: Days
• US Class Current: 435/91.1
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/052 inducing gain of function
• CPC Class Codes: A01K 2217/072 maintaining or altering function, i.e. knock in
• CPC Class Codes: A01K 2217/15 Animals comprising multiple alterations of the genome, by transgenesis or homologous recombination, e.g. obtained by cross-breeding
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 2267/02 Animal zootechnically ameliorated
• CPC Class Codes: A01K 2267/0381 Animal model for diseases of the hematopoietic system
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• 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 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/461 Igs containing Ig-regions, -domains or -residues form different species
• 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 2317/10 characterized by their source of isolation or production
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• 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 2317/50 characterized by immunoglobulin fragments
• CPC Class Codes: C07K 2317/515 : Complete light chain, i.e. VL + CL
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/64 : comprising a combination of variable region and constant region components
• CPC Class Codes: C12N 15/10 : Processes for the isolation, preparation or purification of DNA or RNA chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34
• CPC Class Codes: C12N 15/8509 : for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 5/0606 : Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

Genetically modified mice are provided that express human λ variable (hVλ) sequences, including mice that express hVλ sequences from an endogenous mouse λ light chain locus, mice that express hVλ sequences from an endogenous mouse κ light chain locus, and mice that express hVλ sequences from a transgene or an episome wherein the hVλ sequence is linked to a mouse constant sequence. Mice are provided that are a source of somatically mutated human λ variable sequences useful for making antigen-binding proteins. Compositions and methods for making antigen-binding proteins that comprise human λ variable sequences, including human antibodies, are provided.

Claims

1. A mouse comprising in its germline, at an endogenous mouse light chain locus, a human λ

light chain variable region sequence, wherein the human λ

variable region sequence is expressed in a light chain that comprises a mouse immunoglobulin constant region gene sequence.

2. The mouse of claim 1, wherein the endogenous mouse light chain locus is a λ

locus.

3. The mouse of claim 1, wherein the endogenous mouse light chain locus is a κ

locus.

4. The mouse of claim 1, wherein the mouse lacks an endogenous light chain variable sequence at the endogenous mouse light chain locus.

5. The mouse of claim 1, wherein all or substantially all endogenous mouse light chain variable region gene segments are replaced with one or more human λ

variable region gene segments.

6. The mouse of claim 1, wherein the human λ

light chain variable region sequence comprises a human Jλ

sequence.

7. The mouse of claim 6, wherein the human Jλ

sequence is selected from the group consisting of Jλ

1, Jλ

2, Jλ

3, Jλ

7, and a combination thereof.

8. The mouse of claim 1, wherein the human λ

light chain variable region sequence comprises a fragment of cluster A of the human light chain locus.

9. The mouse of claim 8, wherein the fragment of cluster A of the human λ

light chain locus extends from hVλ

3-27 through hVλ

3-1.

10. The mouse of claim 1, wherein the human λ

light chain variable region sequence comprises a fragment of cluster B of the human light chain locus.

11. The mouse of claim 10, wherein the fragment of cluster B of the human λ

light chain locus extends from hVλ

5-52 through hVλ

1-40.

12. The mouse of claim 1, wherein the human λ

light chain variable region sequence comprises a genomic fragment of cluster A and a genomic fragment of cluster B.

13. The mouse of claim 12, wherein the human λ

light chain variable region sequence comprises at least one gene segment of cluster A and at least one gene segment of cluster B.

14. The mouse of claim 1, wherein more than 10% of the light chain repertoire of the mouse is derived from at least two hVλ

gene segments selected from 2-8, 2-23, 1-40, 5-45, and 9-49.

15. The mouse of claim 14, wherein more than 20% of the light chain repertoire of the mouse is derived from at least three hVλ

gene segments selected from 2-8, 2-23, 1-40, 5-45, and 9-49.

16. The mouse of claim 15, wherein more than 30% of the light chain repertoire of the mouse is derived from at least four hVλ

gene segments selected from 2-8, 2-23, 1-40, 5-45, and 9-49.

17. A mouse that expresses an immunoglobulin light chain that comprises a human lambda variable sequence fused with a mouse constant region, wherein the mouse exhibits a κ

usage to λ

usage ratio of about 1;

1.

18. The mouse of claim 17, wherein the immunoglobulin light chain is expressed from an endogenous mouse light chain locus.

19. A method for making a human λ

variable region nucleic acid sequence, comprising a step of cloning from a cell a human λ

variable region nucleic acid sequence, wherein the cell is derived from a mouse that comprises in its germline a hVλ

gene segment and a hJλ

gene segment operably linked to a mouse constant region nucleic acid sequence at an endogenous mouse constant light chain locus.

20. The method of claim 19, wherein the cell is a hybridoma.