# US 11,997,994 B2 · Genetically modified non-human animals with common light chain immunoglobulin locus

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- Number: US 11,997,994 B2
- Document ID: US11997994B2
- Status: Granted
- Kind: B2
- Application number: 17/969,254
- Filed: 2022-10-19
- Published: 2023-04-27
- Granted: 2024-06-04
- Earliest claimed: 2020-06-02
- Assignees: Biocytogen Pharmaceuticals (Beijing) Co., Ltd.
- Applicants: Biocytogen Pharmaceuticals (Beijing) Co., Ltd.
- Inventors: Yabo Zhang; Hui Lu; Huizhen Zhao; Jiawei Yao; Yi Yang; Yuelei Shen
- Web page: https://patentgrep.com/patent/US11997994B2

## CPC

- A01K 67/0278: Knock-in vertebrates, e.g. humanised vertebrates
- C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
- C07K 16/2881: {against CD71}
- C07K 16/40: against enzymes
- A01K 2207/15: Humanized animals
- A01K 2217/072: maintaining or altering function, i.e. knock in
- A01K 2227/105: Murine
- A01K 2267/01: Animal expressing industrially exogenous proteins
- A01K 2267/02: Animal zootechnically ameliorated
- C07K 2317/21: from primates, e.g. man
- C07K 2317/56: variable (Fv) region, i.e. VH and/or VL
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

This disclosure relates to genetically modified animals and cells with humanized light chain immunoglobulin locus and/or humanized heavy chain immunoglobulin locus. In one aspect, the endogenous light chain immunoglobulin locus comprises a limit number of human IGKV genes and human IGKJ genes.

## Claims

### Claim 1

A genetically-modified mouse whose genome comprises at the endogenous kappa light chain immunoglobulin locus, an exogenous kappa light chain variable region gene sequence,
wherein the exogenous kappa light chain variable region gene sequence comprises only one human IGKV gene and only one human IGKJ gene,
wherein the human IGKV gene and the one human IGKJ gene are operably linked to an endogenous light chain constant domain gene, and the endogenous kappa light chain variable region gene sequence is nonfunctional, wherein
(1) the human IGKV gene is IGKV 1-39 and the human IGKJ gene is IGKJ4; or
(2) the human IGKV gene is IGKV 3-11 and the human IGKJ gene is IGKJ1,
wherein the mouse produces an antibody comprising a human light chain variable region (VL) in response to an antigen.

### Claim 2 (depends on 1)

The mouse of claim 1, wherein the exogenous light chain variable region gene sequence further comprises a human IGKJ 3′-UTR sequence.

### Claim 3 (depends on 1)

The mouse of claim 1, wherein the exogenous light chain variable region gene in one or more cells of the mouse is subject to somatic hypermutations.

### Claim 4 (depends on 1)

The mouse of claim 1, wherein the human IGKV gene is IGKV1-39 and the human IGKJ gene is IGKJ4.

### Claim 5 (depends on 1)

The mouse of claim 1, wherein the human IGKV gene is IGKV3-11 and the human IGKJ gene is IGKJ1.

### Claim 6 (depends on 1)

The mouse of claim 1, wherein the mouse further comprises a promoter sequence that is operably linked to the human IGKV gene, wherein the promoter sequence is within 3000 bp of the human IGKV gene.

### Claim 7 (depends on 1)

The mouse of claim 1, wherein the mouse comprises an endogenous IGKC.

### Claim 8 (depends on 1)

The mouse of claim 1, wherein the mouse is homozygous or heterozygous with respect to the light chain immunoglobulin gene locus.

### Claim 9 (depends on 1)

The mouse of claim 1, wherein the mouse further comprises at an endogenous heavy chain immunoglobulin gene locus, one or more human IGHV genes, one or more human IGHD genes, and one or more human IGHKJ genes, wherein the human IGHV genes, the human IGHD genes, and the human IGHJ genes are operably linked and can undergo VDJ rearrangement.

### Claim 10 (depends on 1)

An isolated cell obtained from the mouse of claim 1.

### Claim 11 (depends on 1)

The mouse of claim 1, wherein the mouse produces an antibody or antigen binding fragment thereof comprising a human light chain variable region that has an amino acid sequence that is at least 90% identical to SEQ ID NO: 38 or 39.

### Claim 12 (depends on 1)

The mouse of claim 1, wherein the mouse produces an antibody or antigen binding fragment thereof comprising a human light chain variable region that comprises the amino acid sequence as set forth in SEQ ID NO: 38.

### Claim 13 (depends on 1)

The mouse of claim 1, wherein the mouse produces an antibody or antigen binding fragment thereof comprising a human light chain variable region that comprises the amino acid sequence as set forth in SEQ ID NO: 39.

### Claim 14 (depends on 1)

The mouse of claim 1, wherein the exogenous light chain variable region gene sequence can be detected by PCR using one or more primer pairs selected from the group consisting of:
(a) SEQ ID NO: 8 and SEQ ID NO: 9;
(b) SEQ ID NO: 10 and SEQ ID NO: 11;
(c) SEQ ID NO: 16 and SEQ ID NO: 17; and
(d) SEQ ID NO: 18 and SEQ ID NO: 19.

### Claim 15 (depends on 1)

The mouse of claim 1, wherein the mouse comprises at the endogenous light chain immunoglobulin locus a nucleic acid sequence that is 95% identical to SEQ ID NO: 35.

### Claim 16 (depends on 1)

The mouse of claim 1, wherein the mouse comprises at the endogenous light chain immunoglobulin locus a nucleic acid sequence that is identical to SEQ ID NO: 35.

### Claim 17 (depends on 1)

The mouse of claim 1, wherein the mouse comprises at the endogenous light chain immunoglobulin locus a nucleic acid sequence that is 95% identical to SEQ ID NO: 36.

### Claim 18 (depends on 1)

The mouse of claim 1, wherein the mouse comprises at the endogenous light chain immunoglobulin locus a nucleic acid sequence that is identical to SEQ ID NO: 36.