Monoclonal antibody that recognizes an epitope of the sperm equatorial region and specifically inhibits sperm-oolemma fusion but not binding.
Balb/c mice were immunized with purified hamster sperm heads for induction of antisera and the production of monoclonal antibodies that recognize preferentially the equatorial segment. Twenty-six hybridoma clones secreted monoclonal antibodies with strong affinity for spermatozoa. The supernatants of 16 clones contained antibodies against the equatorial segment, of which 11 were specific to this region. Five supernatants (M1-M5) containing antibodies that bind to various regions of the sperm head were selected and assessed for the ability to inhibit hamster fertilization in vitro using intact and zona-free oocytes. All the supernatants inhibited fertilization compared with the control. However, M1 supernatant specifically inhibited sperm-egg fusion in a concentration-dependent manner, while sperm-oolemma binding and sperm motility remained unaffected. M1 supernatant recognized an epitope that is exclusive to the equatorial segment and expression of this epitope increased after capacitation and the acrosome reaction. Preliminary immunoblot analysis indicated that M1 monoclonal antibody recognized two protein bands of 37.5 and 34.0 kDa.
Noor
M M
MM
Department of Molecular Biology and Biotechnology, University of Sheffield, UK.
Moore
H D
HD
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D006224:Cricetinae; D004306:Dose-Response Relationship, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006825:Hybridomas; D015151:Immunoblotting; D008297:Male; D008561:Membrane Fusion / Q000276:immunology; D008647:Mesocricetus; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D013077:Sperm Head / Q000276:immunology* / Q000648:ultrastructure; D013084:Sperm-Ovum Interactions / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes