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

Journal Article · Sat Feb 15 04:00:00 EST 1986 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(86)90062-X· OSTI ID:5491367

Pierce, E A; Dame, M C; DeLuca, H F

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 (approx. 100 pM IgG). The RISA has been used successfully to screen for monoclonal antibodies to the intracellular receptor for 1,25-dihydroxyvitamin D/sub 3/ and should be useful for the detection of antibodies to ligand-binding proteins in general.

• Research Organization: Univ. of Wisconsin, Madison
• OSTI ID: 5491367
• Journal Information: Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 153:1; ISSN ANBCA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
ANTIBODIES
ANTIGENS
BODY
CHOLECALCIFEROL
HOMOGENATES
HORMONES
HYBRIDOMAS
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
LIGANDS
MAMMALS
MEMBRANE PROTEINS
MICE
MONOCLONAL ANTIBODIES
ORGANIC COMPOUNDS
PROTEINS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNOLOGY
RECEPTORS
RODENTS
TISSUES
TRACER TECHNIQUES
VERTEBRATES
VITAMIN D
VITAMINS