Gamma ray-induced mutants as a tool for the production and characterisation of monoclonal antibodies against HLA-alloantigens.
To simplify the screening procedure for murine monoclonal antibodies specific for polymorphic HLA determinants, spleen cells from a mouse immunized with the human cell line BJAB-B95.8.6 were fused with NS1 mouse myeloma cells, and hybridoma supernatants were screened for their reactivity on BJAB-B95.8.6 and two gamma ray-induced HLA-loss mutants of this line. The use of these HLA-loss mutants allowed the rapid identification of two new allospecific MOABs designated TU160 and TU161. Serological as well as biochemical studies revealed TU160 to be specific for HLA-A2, and TU161 for HLA-B13 molecules, respectively. Both MOABs were determined to be antibodies of the IgG class and were able to precipitate their antigens from lysates of radioactively labeled cells.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006680:HLA Antigens / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D015789:HLA-A2 Antigen; D015235:HLA-B Antigens; D059908:HLA-B13 Antigen; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000378:metabolism / Q000528:radiation effects; D007126:Immunoglobulin Allotypes / Q000032:analysis / Q000276:immunology; D007074:Immunoglobulin G / Q000032:analysis; D007525:Isoelectric Focusing; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D011233:Precipitin Tests
D000911:Antibodies, Monoclonal; D006680:HLA Antigens; D015789:HLA-A2 Antigen; D015235:HLA-B Antigens; D059908:HLA-B13 Antigen; D007126:Immunoglobulin Allotypes; D007074:Immunoglobulin G