NON-HUMAN MAMMALS FOR THE PRODUCTION OF CHIMERIC ANTIBODIES

• US 20160219848A1
• Filed: 04/11/2016
• Published: 08/04/2016
• Est. Priority Date: 09/30/2008
• Status: Active Grant

• Current Assignee: Ablexis LLC
• Original Assignee: Ablexis LLC
• Inventors: GREEN, Larry, SHIZUYA, Hiroaki
• Granted Patent: US 10,492,476 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A01K 2207/15 Humanized animals
• 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 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 16/283 against Fc-receptors, e.g. CD16, CD32, CD64 CD23 C07K16/2851
• 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 2317/14 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 2317/51 Complete heavy chain or Fd fragment, i.e. VH + CH1
• CPC Class Codes: C07K 2317/515 Complete light chain, i.e. VL + CL
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/522 CH1 domain
• CPC Class Codes: C07K 2317/524 CH2 domain
• CPC Class Codes: C07K 2317/526 CH3 domain
• CPC Class Codes: C07K 2317/53 Hinge
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 : Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: C12N 15/8509 : for producing genetically modified animals, e.g. transgenic

Abstract

The invention provides knock-in non-human cells and mammals having a genome encoding chimeric antibodies and methods of producing knock-in cells and mammals. Certain aspects of the invention include chimeric antibodies, humanized antibodies, pharmaceutical compositions and kits. Certain aspects of the invention also relate to diagnostic and treatment methods using the antibodies of the invention.

Claims

1. A mouse whose genome comprises an immunoglobulin (Ig) lambda light chain locus comprising a transgene comprising all or a portion of a human Igλ

locus, the transgene comprising a DNA fragment comprising in operable linkage from 5′

to 3′

a DNA sequence of a human variable lambda (Vλ

) segment gene operably linked to a constant region gene comprising a human Cλ and

an Igλ

3′

locus control region (LCR), wherein the transgene comprising the human Igλ

locus replaces all or a portion of the endogenous Igλ

locus, thereby the mouse comprises a genome encoding the human Igλ

light chain.

2. The mouse according to claim 1, wherein the portion of the endogenous Igλ

locus comprises one or more of mouse Igλ

3′

LCR, mouse Eλ

2-4, and mouse Eλ

3-1.

3. The mouse according to claim 1, wherein the human Igλ

light chain locus comprises all of a human Igλ

light chain locus.

4. The mouse according to claim 1, wherein the human Igλ

light chain locus comprises one or more human Vλ

segment genes and 1 to 7 human Jλ

-Cλ

segment gene pairs.

5. The mouse according to claim 1, wherein the human Igλ

light chain locus comprises one or more human Vλ

segment genes, 1 to 7 human Jλ

segment genes, and a single human Cλ

segment gene.

6. The mouse according to claim 1, wherein the Igλ

3′

LCR binds nuclear factor kappa-light-chain-enhancer of activated B cells (NFκ

B).

7. The mouse according to claim 6, wherein the Igλ

3′

LCR is human, non-human primate, or rat.

8. The mouse according to claim 7, wherein the Igλ

3′

LCR is human.

9. The mouse according to claim 1, wherein the mouse Igλ

3′

LCR, mouse Eλ

2-4, and mouse Eλ

3-1 are replaced.

10. A mouse cell whose genome comprises an Igλ

locus comprising a transgene comprising all or a portion of a human Igλ

locus, the transgene comprising a DNA fragment comprising in operable linkage from 5′

to 3′

a DNA sequence of a human Vλ

segment gene operably linked to a constant region gene comprising a human Cλ and

an Igλ

3′

LCR, wherein the transgene comprising the human Igλ

locus replaces all or a portion of the endogenous Igλ

locus, thereby the mouse cell comprises a genome encoding the human Igλ

light chain.

11. The mouse cell according to claim 10, wherein the portion of the endogenous Igλ

locus comprises one or more of mouse Igλ

3′

LCR, mouse Eλ

2-4, and mouse Eλ

3-1.

12. The mouse cell according to claim 10, wherein the human Igλ

light chain locus comprises all of a human Igλ

light chain locus.

13. The mouse cell according to claim 10, wherein the human Igλ

light chain locus comprises one or more human Vλ

segment genes and 1 to 7 human Jλ

-Cλ

segment gene pairs.

14. The mouse cell according to claim 10, wherein the human Igλ

light chain locus comprises one or more human Vλ

segment genes, 1 to 7 human Jλ

segment genes, and a single human Cλ

segment gene.

15. The mouse cell according to claim 10, wherein the Igλ

3′

LCR binds NFκ

B.

16. The mouse cell according to claim 15, wherein the Igλ

3′

LCR is human, non-human primate, or rat.

17. The mouse cell according to claim 16, wherein the Igλ

3′

LCR is human.

18. The mouse cell according to claim 10, wherein the mouse Igλ

3′

LCR, mouse Eλ

2-4, and mouse Eλ

3-1 are replaced.

19. The mouse cell according to claim 10, wherein the cell is an embryonic stem cell.

20. The mouse cell according to claim 10, wherein the cell is a B cell.

21. The mouse cell according to claim 10, wherein the cell is a hybridoma cell.