MONOCLONAL-ANTIBODIES REACTING AGAINST CAPACITATED BUT NOT WITH FRESHLY EJACULATED BOAR SPERMATOZOA

PROBLEM: The involvement of individual sperm proteins in differentiati on of antigenically specific and functionally defined regions on sperm membrane has not yet been completely elucidated. METHOD: BALB/c mice were immunized with Live capacitated boar spermatozoa and used for pro duction of monoclonal antibodies (MAbs). ELISA, IIF, SDS-PAGE, IVF, an d cytologic methods were used for selection and biological characteriz ation of MAbs as well as for identification of corresponding antigens. RESULTS: MAb1F10, MAb2E2, and MAb4B12 react with antigens in the acro some portion of live capacitated spermatozoa. MAb1F10 reacted with hum an sperm cells along with those from bull, ram, mouse, dog, whereas MA b2E2-with mouse's spermatozoa and MAB4B12-with bull's, mouse's, and do g's spermatozoa. Some glycolytic enzymes seemed to reduce mildly the r eactions of the MAbs with enzyme treated sperm cells, proteolytic enzy mes eliminated the binding of MAbs to the sperm acrosome. These MAbs h ave no sperm agglutinating and/or sperm-immobilizing activities and re duced the number of spermatozoa binding to zona pellucida. CONCLUSIONS : MAb1F10, MAb2E2, and MAb4B12 seemed to recognize membrane associated antigens with potential role in the initial stages of fertilization, specific for capacitated but not for freshly ejaculated spermatozoa.