Non-human mammals for the production of chimeric antibodies

• US 10,492,476 B2
• Filed: 04/11/2016
• Issued: 12/03/2019
• Est. Priority Date: 09/30/2008
• Status: Active Grant

• Current Assignee: Ablexis LLC
• Original Assignee: Ablexis LLC
• Inventors: Green, Larry, Shizuya, Hiroaki
• Primary Examiner(s): Leavitt, Maria G
• Application Number:
  US15/095,900
• Publication Number:
  US 20160219848A1
• Time in Patent Office: 1,331 Days
• Field of Search: None
• 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 lambda (Igλ

) light chain locus comprising a transgene comprising all or a portion of an unrearranged human Igλ

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

to 3′

a DNA sequence of one or more human variable lambda (Vλ

) gene segments 1 to 7 human Jλ

-Cλ

gene segment pairs and an Igλ

3′

locus control region (LCR), wherein the Igλ

3′

LCR is human, non-human primate, or rat, wherein the transgene comprising the human Igλ

locus replaces all or a portion of the endogenous Igλ

locus, and wherein the replaced portion of the endogenous Igλ

locus comprises mouse Ig lambda Enhancer 2-4 (Eλ

2-4) and/or mouse Ig lambda Enhancer 3-1 (Eλ

3-1), wherein the transgene is capable of undergoing gene rearrangement to produce upon expression a a human Igλ

chain.

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

light chain locus comprises all of a human Igλ

light chain locus.

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

3′

LCR is human.

4. An isolated mouse cell whose genome comprises an Igλ

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

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

to 3′

a DNA sequence of one or more human variable lambda (Vλ

) gene segments 1 to 7 human Jλ

-Cλ

, gene segment pairs and an Igλ

3′

locus control region (LCR), wherein the Igλ

3′

LCR is human, non-human primate, or rat, wherein the transgene comprising the human Igλ

locus replaces all or a portion of the endogenous Igλ

locus, and wherein the replaced portion of the endogenous Igλ

locus comprises mouse Ig lambda Enhancer 2-4 (Eλ

2-4) and/or mouse Ig lambda Enhancer 3-1 (Eλ

3-1), wherein the transgene is capable of undergoing gene rearrangement to produce upon expression a a human Igλ

chain.

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

light chain locus comprises all of a human Igλ

light chain locus.

6. The mouse cell according to claim 4, wherein the Igλ

3′

LCR is human.

7. The mouse cell according to claim 4, wherein the cell is an embryonic stem cell.

8. The mouse cell according to claim 4, wherein the cell is a B cell.

9. The mouse cell according to claim 4, wherein the cell is a hybridoma cell.

10. A mouse whose genome comprises an immunoglobulin lambda (Igλ

) light chain locus comprising a transgene comprising all or a portion of an unrearranged human Igλ

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

to 3′

a DNA sequence of one or more human variable lambda (Vλ

) gene segments 1 to 7 human Jλ

-Cλ

, gene segment pairs and an immunoglobulin kappa (Igκ

) 3′

locus control region (LCR), wherein the Igκ

3′

LCR is human, non-human primate, or rat, wherein the transgene comprising the human Igλ

, locus replaces all or a portion of the endogenous Igλ

, locus, and wherein the replaced portion of the endogenous Igλ

, locus comprises mouse Ig lambda Enhancer 2-4 (Eλ

2-4) and/or mouse Ig lambda Enhancer 3-1 (Eλ

3-1), wherein the transgene is capable of undergoing gene rearrangement to produce upon expression a a human Igλ

chain.