Epitopic analysis of detergent-solubilized HLA molecules by solid-phase radioimmunoassay.
A solid-phase radioimmunoanalysis of plasma membrane molecules solubilized in the presence of detergent was developed. From a crude cell lysate, membrane molecules were specifically immobilized by solid-phase adsorbed monoclonal antibodies. Analysis of these molecules was easily performed with radiolabeled monoclonal antibodies of different specificities. This technique was used to analyze the HLA-DR and HLA-A,B,C molecules expressed by RAJI cells. It was possible (a) to determine which epitopes are expressed on the same molecules; (b) to define subsets of HLA molecules according to the epitopes which they express, and (c) to perform quantitative analysis of HLA molecules on a crude cell lysate.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D006680:HLA Antigens / Q000032:analysis* / Q000276:immunology; D015235:HLA-B Antigens; D015236:HLA-C Antigens; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II / Q000032:analysis / Q000276:immunology; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011863:Radioimmunoassay / Q000379:methods*; D012995:Solubility
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006680:HLA Antigens; D015235:HLA-B Antigens; D015236:HLA-C Antigens; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II; C024062:nonidet; D011092:Polyethylene Glycols