Methods of producing hybrid antibodies
View Patent ↗A method for engineering and utilizing large DNA vectors to target, via homologous recombination, and modify, in any desirable fashion, endogenous genes and chromosomal loci in eukaryotic cells. These large DNA targeting vectors for eukaryotic cells, termed LTVECs, are derived from fragments of cloned genomic DNA larger than those typically used by other approaches intended to perform homologous targeting in eukaryotic cells. Also provided is a rapid and convenient method of detecting eukaryotic cells in which the LTVEC has correctly targeted and modified the desired endogenous gene(s) or chromosomal locus (loci) as well as the use of these cells to generate organisms bearing the genetic modification.
1. A method of producing an antibody, the method comprising:
(a) providing a genetically modified mouse comprising in its germline human unrearranged heavy chain variable region gene segments operably linked to a mouse heavy chain constant region gene at an endogenous mouse heavy chain immunoglobulin locus, wherein rearrangement of the human heavy chain variable region gene segments in the mouse results in a rearranged 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 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 mouse does not produce fully human antibodies;
(b) stimulating an immune response in the genetically modified mouse by exposing the genetically modified mouse to an antigen; and
(c) obtaining the hybrid antibody from the serum of the genetically modified mouse.
2. The method of claim 1 , wherein the mouse heavy chain constant region gene encodes a murine Fc region.
3. A method of producing an antibody, the method comprising:
(a) providing a genetically modified mouse comprising in its germline:
(i) a hybrid heavy chain immunoglobulin locus comprising human unrearranged heavy chain variable region gene segments operably linked to a mouse heavy chain constant region gene at an endogenous mouse heavy chain immunoglobulin locus, wherein rearrangement of the human heavy chain variable region gene segments in the mouse results in a rearranged human heavy chain variable region gene linked to the mouse heavy chain constant region gene; and
(ii) a hybrid light chain immunoglobulin locus comprising human unrearranged light chain variable region gene segments operably linked to a mouse light chain constant region gene at an endogenous mouse light chain immunoglobulin locus, wherein rearrangement of the human light chain variable region gene segments in the mouse results in a rearranged human light chain variable region gene linked to the mouse light chain constant region gene, wherein the mouse in response to an antigen produces a hybrid antibody, the hybrid antibody comprising a heavy chain comprising 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 and a light chain comprising a human light chain variable region encoded by the rearranged human light chain variable region gene and a mouse light chain constant region encoded by the mouse light chain constant region gene; wherein the mouse does not produce fully human antibodies;
(b) stimulating an immune response in the genetically modified mouse by exposing the genetically modified mouse to an antigen; and
(c) obtaining the hybrid antibody from the serum of the genetically modified mouse.
4. The method of claim 3 , wherein the mouse heavy chain constant region gene encodes a murine Fc region.
5. The method of claim 3 , wherein the human light chain variable region gene segments are kappa human light chain gene segments.