A radiometric immunosorbent assay for the detection of anti-hormone-binding protein antibodies.
A radiometric immunosorbent assay (RISA) for the detection of monoclonal antibodies to hormone-binding proteins has been developed. The assay involves incubating hybridoma supernatants in microtiter wells that have been coated with goat anti-mouse IgG antibodies. Any mouse IgG in the test supernatant is thus specifically retained in the wells. Radioactive ligand-binding protein complexes are then incubated in the wells. The presence of anti-binding protein antibodies in the supernatant is indicated by specific retention of radioactive ligand-binding protein complexes in the wells. Crude antigen preparations, such as tissue homogenates, can be used to detect antibodies. The assay is capable of detecting antibody at concentrations 20 ng/ml (approximately 100 pM IgG). The RISA has been used successfully to screen for monoclonal antibodies to the intracellular receptor for 1,25-dihydroxyvitamin D3 and should be useful for the detection of antibodies to ligand-binding proteins in general.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000276:immunology; D000941:Antigens / Q000276:immunology; D006041:Goats; D006801:Humans; D007413:Intestinal Mucosa / Q000276:immunology; D051379:Mice; D011863:Radioimmunoassay / Q000379:methods*; D011864:Radioimmunosorbent Test / Q000379:methods*; D018167:Receptors, Calcitriol; D011956:Receptors, Cell Surface / Q000276:immunology*; D011987:Receptors, Steroid / Q000276:immunology; D013552:Swine; D014809:Vitamin D-Binding Protein / Q000276:immunology
D000911:Antibodies, Monoclonal; D000941:Antigens; D018167:Receptors, Calcitriol; D011956:Receptors, Cell Surface; D011987:Receptors, Steroid; D014809:Vitamin D-Binding Protein