A radiometric immunosorbent assay for the detection of anti-hormone-binding protein antibodies

✍ Scribed by Eric A. Pierce; Margaret C. Dame; Hector F. DeLuca

• Publisher: Elsevier Science
• Year: 1986
• Tongue: English
• Weight: 963 KB
• Volume: 153
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(86)90062-x

✦ Synopsis

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.