Monoclonal antibodies reacting against capacitated but not with freshly ejaculated boar spermatozoa.

PROBLEM

The involvement of individual sperm proteins in differentiation 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 production of monoclonal antibodies (MAbs). ELISA, IIF, SDS-PAGE, IVF, and cytologic methods were used for selection and biological characterization of MAbs as well as for identification of corresponding antigens.

RESULTS

MAb1F10, MAb2E2, and MAb4B12 react with antigens in the acrosome portion of live capacitated spermatozoa, MAb 1F10 reacted with human sperm cells along with those from bull, ram, mouse, dog, whereas MAb2E2--with mouse's spermatozoa and MAB4B12-with bull's, mouse's, and dog's spermatozoa. Some glycolytic enzymes seemed to reduce mildly the reactions of the MAbs with enzyme treated sperm cells; proteolytic enzymes eliminated the binding of MAbs to the sperm acrosome. These MAbs have no sperm agglutinating and/or sperm-immobilizing activities and reduced 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.