PURIFICATION AND CHARACTERIZATION OF SPERM-COATING ANTIGEN, IDENTIFIED BY A MONOCLONAL-ANTIBODY

A procedure was designed for purification of a human seminal plasma-sp ecific antigen (HSP-antigen) identified by a monoclonal antibody produ ced in this laboratory (mAb 4E6). Pooled human seminal plasma was frac tionated by consecutively applied methods: affinity chromatography on Lentil lectin sepharose, gel chromatography on Ultrogel AcA 34 and imm unoaffinity on mAb 4E6 coupled CNBr-Spharose 4B. The antigen-containin g fraction obtained after this procedure was proved to be homogeneous when analysed by electrophoresis in polyacrylamide gel. After the cons ecutive purification procedures the degree of purification was 128 tim es as compared to the starting material. Electrophoretic analysis of t he purified 4E6 antigen under reducing conditions showed that it consi sted of 3 polypeptide subunits with molecular weight 70 kDa, 64 kDa an d 60 kDa respectively. On the basis of the data obtained from competit ive ELISA testing of sera from infertile patients it has been suggeste d that the identified antigen may be involved in pathogenesis of immun ologic infertility.