Patent ID: 8293480
Filing Date: 2012-10-23
Classification: A01K,C07K,C12N

Abstract:
1. A method for identifying a murine IgG isotype immunoglobulin molecule comprising a human variable region that specifically binds to a predetermined antigen, wherein the immunoglobulin molecule is expressed by a B lymphocyte hybridoma clone, wherein the B lymphocyte hybridoma clone is derived from a fusion of: (i) a B lymphocyte from a transgenic mouse, wherein the genome of said transgenic mouse comprises inactivated murine variable regions and an unrearranged human heavy chain immunoglobulin variable region operably linked to a mu constant region gene segment, wherein the unrearranged heavy chain variable region comprises multiple human VH gene segments, multiple human D gene segments, and multiple human JH gene segments, and wherein the mu constant region is selected from the group consisting of: (ii) a myeloma cell; and which method comprises: (i) incubating a first antibody mixture derived from the B lymphocyte hybridoma clone containing the murine IgG isotype immunoglobulin molecule expressed by said B lymphocyte hybridoma clone with said antigen under conditions sufficient to allow for specific binding, (ii) modifying the incubation conditions to remove non-specifically bound antibodies, (iii) incubating a second antibody mixture containing an anti-murine IgG antibody comprising a marker under conditions sufficient to allow for specific binding of the anti-murine IgG antibody to murine IgG antibody, (iv) modifying the incubation conditions to remove non-specifically bound anti-murine IgG antibodies, and (v) identifying the murine IgG immunoglobulin molecule comprising a human variable region which specifically binds to the predetermined antigen by identifying the presence of the marker.