Mice that produce hybrid antibodies

• US 10,640,800 B2
• Filed: 07/19/2016
• Issued: 05/05/2020
• Est. Priority Date: 02/16/2001
• Status: Expired due to Term

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: Regeneron Pharmaceuticals Incorporated
• Inventors: Murphy, Andrew J., Yancopoulos, George D., Karow, Margaret, MacDonald, Lynn, Stevens, Sean, Economides, Aris N., Valenzuela, David M.
• Primary Examiner(s): Singh, Anoop K
• Assistant Examiner(s): Sgagias, Magdalene K
• Application Number:
  US15/213,947
• Publication Number:
  US 20160316731A1
• Time in Patent Office: 1,386 Days
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• 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 16/28 against receptors, cell surface antigens or cell surface determinants
• 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 2317/10 characterized by their source of isolation or production
• 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 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C12N 15/67 General methods for enhancing the expression
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 15/902 using homologous recombination
• CPC Class Codes: C12N 15/907 in mammalian cells
• CPC Class Codes: C12N 2800/204 : of bacterial origin, e.g. BAC
• CPC Class Codes: C12P 21/00 : Preparation of peptides or proteins single cell protein C12N1/00

Abstract

Provided herein is a mouse that produces hybrid antibodies containing human variable regions and mouse constant regions.

Claims

1. A mouse comprising in its germline unrearranged human heavy chain variable region gene segments, wherein the unrearranged human heavy chain variable region gene segments in situ replace endogenous mouse heavy chain immunoglobulin variable region gene segments, and the unrearranged human heavy chain variable region gene segments are operably linked to an endogenous mouse heavy chain constant region gene, wherein rearrangement of the unrearranged human heavy chain variable region gene segments in the mouse results in a rearranged human heavy chain variable region gene operably linked to the mouse heavy chain constant region gene, wherein in response to exposure to an antigen the mouse produces a hybrid antibody to the antigen, wherein the hybrid antibody comprises a human heavy chain variable region encoded by the rearranged human heavy chain variable region gene and a mouse heavy chain constant region encoded by the mouse heavy chain constant region gene, and wherein the endogenous immunoglobulin enhancer Eμ

remains intact upstream of the mouse heavy chain constant region gene.

2. The mouse of claim 1, wherein the human heavy chain variable region gene segments are contained on a human genomic DNA fragment that is larger than 20 kb.

3. The mouse of claim 1, wherein the human heavy chain variable region gene segments are contained on a human genomic DNA fragment that is larger than 100 kb.

4. The mouse of claim 1, wherein the mouse further comprises in its germline unrearranged human light chain variable region gene segments operably linked to a mouse light chain constant region gene.

5. The mouse of claim 4, wherein the human heavy chain variable region gene segments are contained on a human genomic DNA fragment that is larger than 20 kb and the human light chain variable region gene segments are contained on a human genomic DNA fragment that is larger than 20 kb.

6. The mouse of claim 4, wherein the human heavy chain variable region gene segments are contained on a human genomic DNA fragment that is larger than 100 kb and the human light chain variable region gene segments are contained on a human genomic DNA fragment that is larger than 100 kb.