Human lambda light chain mice

• US 9,012,717 B2
• Filed: 06/22/2011
• Issued: 04/21/2015
• 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.
• Primary Examiner(s): WILSON, MICHAEL C
• Application Number:
  US13/166,171
• Publication Number:
  US 20120070861A1
• Time in Patent Office: 1,399 Days
• Field of Search: 800/18
• US Class Current: 800/18
• 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 transgenic mouse whose genome comprises an endogenous λ

light chain immunoglobulin locus comprising;

i) a deletion of a first endogenous Vλ

-Jλ

-Cλ

gene cluster; and

ii) a replacement of a fragment of endogenous Vλ and

Jλ

gene segments in a second endogenous Vλ

-Jλ

-Cλ

gene cluster with human Vλ and

Jλ

gene segments, wherein the human Vλ and

Jλ

gene segments comprise at least one human Vλ

gene segment and at least one human Jλ

gene segment, and wherein the human Vλ and

Jλ

gene segments are operably linked to a mouse Cλ

gene.

2. The transgenic mouse of claim 1, wherein the human Vλ

and Jλ

gene segments comprise a human Jλ

1 gene segment.

3. The transgenic mouse of claim 1, wherein the human Vλ

and Jλ

gene segments comprise four human Jλ

gene segments.

4. The transgenic mouse of claim 3, wherein the four human Jλ

gene segments are Jλ

1, Jλ

2, Jλ

3 and Jλ

7.

5. The transgenic mouse of claim 1, wherein the endogenous mouse Vλ

gene segments of the second endogenous Vλ

-Jλ

-Cλ

gene cluster have been replaced with a contiguous sequence of the human immunoglobulin λ

light chain locus that spans from a human Vλ

3-12 gene segment to a human Vλ

3-1 gene segment.

6. The transgenic mouse of claim 1, wherein the replacement endogenous mouse Vλ

gene segments of the second endogenous Vλ

-Jλ

-Cλ

gene cluster have been replaced with a contiguous sequence of the human immunoglobulin λ

light chain locus that spans from a human Vλ

3-27 gene segment to a human Vλ

3-1 gene segment.

7. The transgenic mouse of claim 1, wherein the replacement comprises endogenous mouse Vλ

gene segments of the second endogenous Vλ

-Jλ

-Cλ

gene cluster have been replaced with a contiguous sequence of the human immunoglobulin λ

light chain locus that spans from a human Vλ

3-27 gene segment to a human Vλ

3-1 gene segment and a contiguous sequence of the human immunoglobulin λ

light chain locus that spans from a human Vλ

5-52 gene segment to a human Vλ

1-40 gene segment.

8. The transgenic mouse of claim 1, wherein the human Vλ

and Jλ

gene segments comprise at least 12 human Vλ

gene segments.

9. The transgenic mouse of claim 8, wherein the 12 human Vλ

gene segments include human Vλ

3-1, Vλ

4-3, Vλ

2-8, Vλ

3-9, Vλ

3-10, Vλ

2-11 and Vλ

3-12.

10. The transgenic mouse of claim 1, wherein the human Vλ

and Jλ

gene segments comprise at least 28 human Vλ

gene segments.

11. The transgenic mouse of claim 10, wherein the 28 human Vλ

gene segments include human Vλ

2-14, Vλ

3-16, Vλ

2-18, Vλ

3-19, Vλ

3-21, Vλ

3-22, Vλ

2-23, Vλ

3-25 and Vλ

3-27.

12. The transgenic mouse of claim 1, wherein the human Vλ

and Jλ

gene segments comprise at least 40 human Vλ

gene segments.

13. The transgenic mouse of claim 12, wherein the 40 human Vλ

gene segments include human Vλ

5-52, Vλ

1-51, Vλ

9-49, Vλ

1-47, Vλ

7-46, Vλ

5-45, Vλ

1-44, Vλ

1-44, Vλ

7-43 and Vλ

1-40.