Patent ID: 9708635
Date: 2017-07-18
CPC Classifications: A01K,C07K,C12N,C12P

Claim:
1. A method of producing a nucleic acid encoding a human immunoglobulin heavy chain variable region, the method comprising: (a) providing a transgenic mouse whose genome comprises in its germline unrearranged human immunoglobulin heavy chain V, D, and J gene segments, wherein the unrearranged human immunoglobulin heavy chain V, D, and J gene segments in situ replace endogenous mouse immunoglobulin heavy chain V, D, and J gene segments, and the unrearranged human immunoglobulin heavy chain V, D, and J gene segments are operably linked to an endogenous mouse heavy chain constant region gene, wherein the mouse heavy chain constant region gene is located at an endogenous mouse immunoglobulin heavy chain constant region locus, wherein rearrangement of the unrearranged human immunoglobulin heavy chain V, D, and J gene segments in the mouse results in a rearranged immunoglobulin human heavy chain variable region gene linked to the mouse heavy chain constant region gene, wherein the mouse in response to an antigen produces a hybrid antibody that comprises a human immunoglobulin heavy chain variable region encoded by the rearranged human immunoglobulin heavy chain variable region gene and a mouse heavy chain constant region encoded by the mouse heavy chain constant region gene; (b) stimulating an immune response in the mouse by exposing the mouse to an antigen; (c) preparing a hybridoma expressing the hybrid antibody from the spleen of the mouse stimulated by the antigen in step (b); and (d) isolating DNA encoding the human immunoglobulin heavy chain variable region of the hybrid antibody from the hybridoma of step (c).