GENETIC ENGINEERING OF NON-HUMAN ANIMALS FOR THE PRODUCTION OF CHIMERIC ANTIBODIES

• US 20170190797A1
• Filed: 03/20/2017
• Published: 07/06/2017
• Est. Priority Date: 03/31/2010
• Status: Active Grant

• Current Assignee: Ablexis LLC
• Original Assignee: Ablexis LLC
• Inventors: Green, Larry, Shizuya, Hiroaki
• Granted Patent: US 10,526,420 B2
• Time in Patent Office: Days
• 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 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• 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 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/461 Igs containing Ig-regions, -domains or -residues form different species
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• 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 2317/64 : comprising a combination of variable region and constant region components
• CPC Class Codes: C12N 15/8509 : for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2510/02 : Cells for production

Abstract

The invention provides non-human cells and mammals having a genome encoding chimeric antibodies and methods of producing transgenic 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 synthetic polynucleotide comprising a gene segment encoding an immunoglobulin heavy chain variable (V) polypeptide, a gene segment encoding an immunoglobulin heavy chain diversity (D) polypeptide, a gene segment encoding an immunoglobulin heavy chain joining (J) polypeptide, and an immunoglobulin heavy chain non-coding element for the V gene segment, wherein the V polypeptide and the J polypeptide are from a first species, wherein the non-coding element is from a second species, and wherein the V gene segment, D gene segment, and J gene segment replace an immunoglobulin heavy chain V coding sequence, an immunoglobulin heavy chain D coding sequence, and an immunoglobulin heavy chain J coding sequence, respectively, of the second species.

2. The synthetic polynucleotide of claim 1, wherein the first species is human.

3. The synthetic polynucleotide of claim 2, wherein the second species is a non-human mammal.

4. The synthetic polynucleotide of claim 3, wherein the second species is mouse.

5. The synthetic polynucleotide of claim 1, wherein the non-coding element is selected from an intron and a cis regulatory element.

6. The synthetic polynucleotide of claim 5, wherein the cis regulatory element is selected from a promoter, an enhancer, a recombination signal sequence and a 5′

untranslated region.

7. The synthetic polynucleotide of claim 1, wherein the V gene segment, the D gene segment, and the J gene segment are in germline configuration.

8. The synthetic polynucleotide of claim 1, wherein the V gene segment, the D gene segment, and the J gene segment are capable of undergoing rearrangement to produce a polynucleotide sequence encoding a polypeptide comprising an immunoglobulin heavy chain variable region.

9. The synthetic polynucleotide of claim 1, wherein the non-coding element and the V polypeptide are from non-orthologous immunoglobulin loci.

10. The synthetic polynucleotide of claim 9, wherein the non-coding element is from an immunoglobulin light chain locus.

11. The synthetic polynucleotide of claim 10, wherein the immunoglobulin light chain locus is a kappa light chain or a lambda light chain.

12. The synthetic polynucleotide of claim 1, further comprising one or more gene segments encoding an immunoglobulin constant (C) polypeptide.

13. The synthetic polynucleotide of claim 12, comprising two or more gene segments encoding two or more C polypeptides, and wherein one of the two or more C polypeptides is from the first species and one of the two or more C polypeptides is from the second species.

14. The synthetic polynucleotide of claim 13, wherein the first species C polypeptide is a CH1 and the second species C polypeptide is a CH2.

15. The synthetic polynucleotide of claim 14, wherein the two or more immunoglobulin C polypeptides are from the second species.

16. The synthetic polynucleotide of claim 1, wherein the non-coding element is an intron or a cis regulatory element.

17. A non-human cell whose genome comprises a transgene comprising the synthetic polynucleotide of claim 1.

18. A non-human mammal whose genome comprises a transgene comprising the synthetic polynucleotide of claim 1.