人精子制动单抗的制备、鉴定及其腹水型单抗的纯化

Preparation,characterization of sperm immobilizing monoclonal antibody and purification from mouse ascites by hydroxyapatite chromatography

用杂交瘤技术建立的一株分泌抗人精浆蛋白的使人精子制动单抗的细胞株，Ｉｇ亚类分析表明，此单抗为ＩｇＭ。间接ＥＬＩＳＡ试验、精子制动试验、精子凝集试验和间接免疫荧光等试验分别证明所得单抗能与人精浆和无精子症患者的精浆产生特异免疫反应，有制动人精子的作用，并能特异结合到人精子的颈部，从而证实此单抗是精子制动单抗，其相应的抗原系精浆和精子共有的精子膜抗原，用ＦＰＬＣ－羟基磷灰石柱层析成功地从小鼠腹水纯化了此单抗。

bjectives:To prepare and to characterize a sperm immobilizing monoclonal antibody against humanseminal plasma and spermatozoa and to purify it from mouse ascitic fluid· Design: An immunological study with human semen. Setting: Laboratory in Zhejiang Medical University·Methods:sperm immobilizing monoclonal antibody was prepared with hybridoma technic· ELISA,sperm immobilization,sperm agglutination, indirect immunofluorescense were tested for the antibody· Hy-droxyapatite chromatography was used for purification of monoclonal antibody·Material: Human seminal plasma protein and sperms were collected from fresh semen samples. BALB/c mouse was used for experiments.Results:The monoclonal antibody prepared was proved to be an lgM according to the analysis of lgsubgroups . It was shown that the monoclonal antibody reacted specifically with seminal plasma of normal and =azospermic men. The antibody was able to immobilize spermatozoa and was bound specifically to theneck of the sperm. The monoclonal antibody was further purified from the mouse ascites with hydroxyapatitechromatography. conclusion:The monoclonal antibody was proved to have an immobilizing effect on human sperms.The responding antigen from the seminal plasma and spermatozoa was a sperm coating antigen.