Isolation of HLA and tumor antigens by means of affinity chromatography employing anti-beta2-microglobulin (beta2m) antiserum.
A method for the isolation of HLA antigen molecules from normal and cancerous solid human tissue is described. The method employs anti-beta2-microglobulin (beta2m) antiserum coupled to Sepharose beads as an immunosorbent affinity medium. The anti-beta2m affinity chromatography procedure greatly purifies and selectively enriches HLA and any material that copurifies by affinity, with beta2m and/or HLA molecules. The HLA isolated by this purification procedure was used to immunize rabbits. The antisera obtained were absorbed on beta2m to remove all anti-beta2m antibody activity. The use of such anti-HLA antisera in radioimmunoassays, immunoprecipitation studies, and F(ab')2 blocking experiments demonstrated that these antisera are directed against a common HLA determinant present on the heavy (alloantigen-bearing) chain of all HLA molecules. The use of an identical procedure employing human tumor tissues has resulted in the isolation of HLA-like or HLA-associated tumor-specific antigens as demonstrated by the leukocyte adherence inhibition (LAI) assay.
D000951:Antigens, Neoplasm / Q000032:analysis*; D001609:Beta-Globulins / Q000276:immunology*; D002846:Chromatography, Affinity / Q000379:methods*; D003602:Cytotoxicity, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D006680:HLA Antigens / Q000032:analysis*; D006801:Humans; D007106:Immune Sera; D007140:Immunoglobulin Fab Fragments; D007957:Leukocyte Adherence Inhibition Test; D008024:Ligands; D009369:Neoplasms / Q000032:analysis*; D011863:Radioimmunoassay; D001613:beta 2-Microglobulin / Q000276:immunology*
D000951:Antigens, Neoplasm; D001609:Beta-Globulins; D006680:HLA Antigens; D007106:Immune Sera; D007140:Immunoglobulin Fab Fragments; D008024:Ligands; D001613:beta 2-Microglobulin