A biotin-streptavidin enzyme immunoassay for detection of antibodies to porcine granulosa cell antigens.
A colorimetric solid-phase enzyme immunoassay has been developed which quantifies antibodies to porcine granulosa cell membrane antigens in rabbits immunized with porcine granulosa cells. A cell-free, particulate membrane preparation of porcine granulosa cells was used as coating antigen. A biotinylated second antibody in conjunction with a streptavidin-beta-galactosidase conjugate was utilized to amplify reactivity. The enzyme beta-galactosidase was used due to high background obtained using peroxidase, presumably due to endogenous peroxidase activity of the tissue. Sigmoidal serum dilution curves were obtained with immune rabbit sera indicating that absorbance was related to the concentration of antibodies. Assay activity was reduced by preincubation of immune serum with granulosa cell membranes. Sera from ovariectomized or pre-immune rabbits did not yield any specific binding in the assay. This assay has potential applicability for quantifying antiovarian and antigranulosa cell antibodies in women suspected of having autoimmune premature ovarian failure.
Kim
J G
JG
Department of Obstetrics and Gynecology, University of Cincinnati College of Medicine, Ohio 45267.
Anderson
B E
BE
Rebar
R W
RW
LaBarbera
A R
AR
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000954:Antigens, Surface / Q000276:immunology*; D001327:Autoimmune Diseases / Q000175:diagnosis*; D001426:Bacterial Proteins; D001710:Biotin; D003124:Colorimetry; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006107:Granulosa Cells / Q000276:immunology*; D006801:Humans; D010052:Ovariectomy; D011107:Polylysine / Q000494:pharmacology; D016649:Primary Ovarian Insufficiency / Q000175:diagnosis*; D011817:Rabbits / Q000276:immunology; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D019809:Streptavidin; D013552:Swine / Q000276:immunology*; D001616:beta-Galactosidase
D000906:Antibodies; D000954:Antigens, Surface; D001426:Bacterial Proteins; D011107:Polylysine; D012710:Serum Albumin, Bovine; D001710:Biotin; D019809:Streptavidin; D001616:beta-Galactosidase