Granted Patent B2

US 12,207,628 · App. 17/129,706 · Granted Jan 28, 2025

Inventors: Lynn Macdonald (Harrison, NY); Sean Stevens (Del Mar, CA); Andrew J. Murphy (Croton-on-Hudson, NY); Margaret Karow (Santa Rosa Valley, CA)

Assignee: Regeneron Pharmaceuticals, Inc.

A01K67/0278A01K67/0275B60H1/32281C07K16/22C07K16/28C07K16/2866C07K16/40C07K16/461C07K16/462C12N9/6489C12N15/8509A01K2207/15A01K2217/072A01K2217/15A01K2227/105A01K2267/01C07K2317/21C07K2317/92C12N2800/204C12N2800/30

Abstract

Claims (26)

1. A method of making a fully human heavy chain or a fully human light chain comprising the steps of:

(a) immunizing a genetically modified mouse with an antigen, wherein the mouse:

(i) has a genome that comprises:

(A) one or more human V H gene segments, one or more human D H gene segments, and one or more human J H gene segments operably linked to a mouse heavy chain constant region at an endogenous mouse immunoglobulin heavy chain locus,

(B) one or more human V L gene segments and one or more human J L gene segments operably linked to a light chain constant region, and

(C) a nucleotide sequence that encodes a mouse ADAM6 protein, which nucleotide sequence is integrated in the genome of the mouse, wherein the mouse ADAM6 protein is expressed in the genetically modified mouse;

(ii) generates antibodies when immunized with the antigen, wherein the antibodies each comprise a human heavy chain variable domain operably linked to a mouse heavy chain constant domain; and

(iii) is fertile;

(b) allowing the genetically modified mouse to mount an immune response to the antigen;

(c) isolating a B cell from the genetically modified mouse that expresses an antibody that specifically binds the antigen;

(d) determining an amino acid sequence of a human heavy or light chain variable domain of an antibody that specifically binds the antigen and that was generated by the genetically modified mouse; and

(e) operably linking the human heavy or light chain variable domain to a human heavy or light chain constant domain, respectively, to form a fully human heavy chain or a fully human light chain.

2. The method of claim 1 , further comprising determining the amino acid sequence of the fully human heavy chain or a fully human light chain.

3. The method of claim 1 , wherein operably linking the human heavy or light chain variable domain to the human heavy or light chain constant domain, respectively, comprises operably linking a nucleotide sequence encoding the human heavy or light chain variable domain to a nucleotide sequence encoding the human heavy or light chain constant domain, respectively.

4. The method of claim 1 , wherein the one or more human V H gene segments comprise a V H 6-1, V H 1-2, V H 1-3, V H 2-5, V H 3-7, V H 1-8, V H 3-9, V H 3-11, V H 3-13, V H 3-15, V H 3-16, V H 1-18, V H 3-20, V H 3-21, V H 3-23, V H 1-24, V H 2-26, V H 4-28, V H 3-30, V H 4-31, V H 3-33, V H 4-34, V H 3-35, V H 3-38, V H 4-39, V H 3-43, V H 1-45, V H 1-46, V H 3-48, V H 3-49, V H 5-51, V H 3-53, V H 1-58, V H 4-59, V H 4-61, V H 3-64, V H 3-66, V H 1-69, V H 2-70, V H 3-72, V H 3-73, V H 3-74, or any combination thereof.

5. The method of claim 1 , wherein the one or more human VL gene segments and one or more human JL gene segments are operably linked to a mouse light chain constant region at an endogenous mouse light chain locus.

6. The method of claim 1 , wherein the one or more human VL gene segments comprise a V κ 1-5, V κ 1-6, V κ 1-8, V κ 1-9, V κ 1-12, V κ 1-16, V κ 1-17, V κ 1-27, V κ 1-33, V κ 1-37, V κ 1-39, V κ 2-24, V κ 2-28, V κ 2-29, V κ 2-30, V κ 2-40, V κ 3-11, V κ 3-15, V κ 3-20, V κ 4-1, V κ 6-21, or any combination thereof.

7. The method of claim 1 , wherein the nucleotide sequence encoding the mouse ADAM6 protein:

(a) is present on the same chromosome as the endogenous mouse immunoglobulin heavy chain locus; and/or

(b) is present at an ectopic position.

8. The method of claim 1 , wherein the mouse ADAM6 protein is a mouse ADAM6a protein or a mouse ADAM6b protein.

9. The method of claim 1 , wherein the nucleotide sequence that encodes the mouse ADAM6 protein is contiguous with one or more human V H gene segments, one or more human D H gene segments, or one or more human J H gene segments.

10. The method of claim 1 , wherein the one or more human V L gene segments are one or more human V κ gene segments and the one or more human J L gene segments are one or more human J κ gene segments.

11. The method of claim 10 , wherein the one or more human V κ gene segments and one or more human J κ gene segments are operably linked to a mouse κ light chain constant region at an endogenous mouse κ light chain locus.

12. The method of claim 1 , wherein the one or more human V L gene segments are one or more human Vλ gene segments and the one or more human J L gene segments are one or more human Jλ gene segments.

13. The method of claim 12 , wherein the one or more human Vλ gene segments and one or more human Jλ gene segments are operably linked to a mouse λ light chain constant region at an endogenous mouse λ light chain locus.

Assignments (1)

Continuity (9)

References Cited (296)