Development of monoclonal antibody for human sperm protein

我们证明了从免疫不孕症妇女获得的血清诱导人类精子头对头凝集，并与人类精子蛋白相互作用。从人类身上收集足量的精液是一项艰巨的任务。为了解决这一供应问题，我们决定从千叶县和福冈县的牲畜实验站和国立畜牧产业研究所研究供应充足的野猪精子。血清还能诱导猪精的头对头凝集。用间接免疫荧光法测定患者制备的精子头顶体区免疫球蛋白G (IgG)，从附睾体和尾部获得。IgG与猪精子蛋白相互作用，SDS-PAGE免疫印迹技术估计分子量为45 kda。然而，IgG不与从睾丸和附睾头或非性腺组织中提取的精子蛋白相互作用。IgG与附睾体液和尾液中存在的约75- kda和38-kDa的附加蛋白相互作用，但与附睾头液中的蛋白不相互作用。相互作用的蛋白在刀豆蛋白A Sepharose色谱柱上被吸附，表明它们是糖蛋白。研究猪精子抗原作为免疫原对实验动物生育能力的影响是研制抗生育疫苗的重要途径。我们已经开发了抗猪精子蛋白的单克隆抗体，在体外诱导猪精子凝集。

We demonstrated that serum obtained from an immunologic infertile woman induced head-to-head agglutination of human sperm and interacted with human sperm proteins. To collect sufficient amounts of semen from human beings is a difficult task. to circumvent this problem of supply, we decided to study boar sperm which is in abundant supply from the Livestock Experimental Station of Chiba and Fukuoka Prefecture, and the National Institute of Animal Industry. The serum also induced a head-to-head agglutination of ejaculated boar sperm. The immunoglobulin G (IgG) prepared from the patient localized to the acrosomal region of the sperm head obatined from the corpus and cauda epididymis as determined by an indirect immunofluorescent method. The IgG interacted with a boar sperm protein with an estimated molecular weight of 45-kDa, determined by SDS-PAGE immunoblotting technique. However, the IgG did not interact with proteins extracted from sperm obtained from the testis and caput epididymis or from non-gonadal tissues. The IgG interacted with additional proteins of about 75- and 38-kDa present in the corpus and cauda epididymal fluids but not those in the caput epididymal fluid. The interacting proteins were adsorbed when chromatographed on Concanavalin A Sepharose column, suggesting that they are glycoproteins.A promising approach in the development of an antifertility vaccine is to study the effects of the boar sperm antigens as immunogens on the fertility of experimental animals. We have developed monoclonal antibodies against boar sperm proteins that induce boar sperm agglutination in vitro.