Solid-phase indirect immunogold localization of human sperm antigens reacting with antisperm antibodies in human sera.
A simple solid-phase indirect immunogold assay for light microscopic localization of the binding of sperm-reactive antibodies in human sera to surface-associated antigenic sites on human sperm is described. Antigen-antibody interaction is localized by using a gold-tagged second antibody directed against the first antibody. Silver enhancement of the 5 nm gold particles makes the immunologic reaction visible by light microscopy. An ELISA was used to identify serum samples as positive or negative for sperm-reactive antibodies. These serum samples were blind-coded and were used to develop and to evaluate this solid-phase indirect immunogold assay. When results obtained by this immunogold assay for the detection of sperm-reactive antibodies were compared with those obtained by the ELISA, there was agreement in 82 of the 100 sera tested. As a localization assay, this indirect immunogold assay is simple to perform and enables stable, high resolution light microscopic localization of the pattern of antigenic sites on sperm surface membranes.
Smithwick
E B
EB
Department of Physiology, Emory University School of Medicine, Atlanta, Georgia.
Young
L G
LG
D000954:Antigens, Surface / Q000032:analysis / Q000276:immunology; D001323:Autoantibodies / Q000097:blood*; D001324:Autoantigens / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D015203:Reproducibility of Results; D013094:Spermatozoa / Q000276:immunology*
D000954:Antigens, Surface; D001323:Autoantibodies; D001324:Autoantigens