NON-HUMAN MAMMALS FOR THE PRODUCTION OF CHIMERIC ANTIBODIES

• US 20160222093A1
• Filed: 04/11/2016
• Published: 08/04/2016
• Est. Priority Date: 09/30/2008
• Status: Active Application

• Current Assignee: Ablexis LLC
• Original Assignee: Ablexis LLC
• Inventors: GREEN, Larry, SHIZUYA, Hiroaki
• Application Number:
  US15/095,889
• Publication Number:
  US 20160222093A1
• 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 non-human mammal whose genome comprises an immunoglobulin (Ig) heavy chain locus comprising a transgene comprising a portion of a syngeneic Ig heavy chain locus, the portion comprising all or part of constant region genes encoding a chimeric Ig heavy chain, the transgene comprising a DNA fragment comprising in operable linkage from 5′

to 3′

a DNA sequence of a human variable heavy (VH) segment gene operably linked to a constant region gene comprising a human CH1, syngeneic mammalian non-human CH2 and syngeneic mammalian non-human CH3 exons, wherein the human CH1 exon is selected from the group consisting of a human mu constant region (Cμ

), a human delta constant region (Cδ

), a human gamma-1 constant region (Cγ

1), a human gamma-2 constant region (Cγ

2), a human gamma-4 constant region (Cγ

4), a human alpha constant region (Cα

), and a human epsilon constant region (Cε

), thereby the non-human mammal comprises a genome encoding the chimeric Ig heavy chain.

2. The non-human mammal according to claim 1, wherein the transgene further comprises a diversity heavy (DH) segment gene 3′

of the human VH segment gene and 5′

of the human CH1 segment gene, wherein the DH segment gene is selected from the group consisting of human, non-human primate, rabbit, sheep, rat, hamster, and mouse.

3. The non-human mammal according to claim 2, wherein the DH segment gene is human.

4. The non-human mammal according to claim 1, wherein the transgene further comprises a human joining heavy (JH) segment gene 3′

of the human VH segment gene and 5′

of the human CH1 segment gene.

5. The non-human mammal according to claim 1, wherein the transgene further comprises a human C_{upper hinge} exon 3′

of the human CH1 exon and 5′

of the non-human CH2 exon.

6. The non-human mammal according to claim 5, wherein the transgene further comprises a human C_{middle hinge} exon 3′

of the human C_{upper hinge} exon and 5′

of the non-human CH2 exon.

7. The non-human mammal according to claim 1, wherein the chimeric Ig heavy chain binds an endogenous Fc receptor.

8. The non-human mammal according to claim 1, wherein the mammal is a mouse.

9. The non-human mammal according to claim 1, wherein the portion further comprises a 3′

locus control region (LCR).

10. The non-human mammal according to claim 1, wherein the transgene comprising the human VH segment gene and the syngeneic Ig heavy chain locus replaces all or a portion of the endogenous Ig heavy chain locus.

11. The non-human mammal according to claim 1, wherein the genome further comprises a transgene comprising a human Ig light chain locus, or a portion thereof.

12. The non-human mammal according to claim 11, wherein the human Ig light chain locus replaces all or a portion of an endogenous Ig light chain locus.

13. The non-human mammal according to claim 12, wherein the human Ig light chain locus comprises a human Igκ

variable region.

14. The non-human mammal according to claim 13, wherein the human Ig light chain locus further comprises a human Igκ

constant region.

15. The non-human mammal according to claim 14, wherein the human Ig light chain locus further comprises a human Igκ

3′

LCR.

16. The non-human mammal according to claim 11, wherein the human Ig light chain locus comprises a human Igλ

variable region.

17. The non-human mammal according to claim 16, wherein the human Ig light chain locus further comprises a human Igλ

constant region.

18. The non-human mammal according to claim 17, wherein the human Ig light chain locus further comprises an Igλ

3′

LCR.

19. The non-human mammal according to claim 17, wherein the human Ig light chain locus comprises all of a human Igλ

light chain locus.

20. The non-human mammal according to claim 17, wherein the human Ig light chain locus comprises one or more human Vλ

segment genes and 1 to 7 human Jλ

-Cλ

segment gene pairs.

21. The non-human mammal according to claim 17, wherein the human Ig light chain locus comprises one or more Vλ

segment genes, 1 to 7 human Jλ

segment genes, and a single human Cλ

segment gene.

22. The non-human mammal according to claim 18, wherein the Igλ

3′

LCR is from a mammal selected from the group consisting of human, non-human primate, and rat.

23. The non-human mammal according to claim 18, wherein the Igλ

3′

LCR is from a mouse and has been engineered to increase NFκ

B binding.