Development of monoclonal antibody for human sperm protein

人精子蛋白单克隆抗体的研制

基本信息

批准号：
05671300
负责人：
HANEJI Tatsuji
金额：
$ 1.41万
依托单位：
Kyushu Dental College
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

项目摘要

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.

我们证明，从免疫性不育妇女获得的血清可诱导人类精子的头对头凝集，并与人类精子蛋白相互作用。从人类身上收集足够量的精液是一项艰巨的任务。为了解决这个供应问题，我们决定研究公猪精子，千叶县和福冈县的畜牧实验站以及国立动物工业研究所供应充足。血清还诱导射出的公猪精子发生头对头凝集。通过间接免疫荧光法测定，从附睾体和附睾尾获得的患者制备的免疫球蛋白G（IgG）位于精子头部的顶体区域。 IgG 与公猪精子蛋白相互作用，通过 SDS-PAGE 免疫印迹技术测定，估计分子量为 45 kDa。然而，IgG 不会与从睾丸和附睾头或非性腺组织获得的精子中提取的蛋白质相互作用。 IgG 与附睾体和尾部液体中存在的约 75 kDa 和 38 kDa 的其他蛋白质相互作用，但与头附睾液中的蛋白质不相互作用。当在伴刀豆球蛋白 A 琼脂糖柱上进行色谱分析时，相互作用的蛋白质被吸附，表明它们是糖蛋白。开发抗生育疫苗的一个有前景的方法是研究公猪精子抗原作为免疫原对实验动物生育能力的影响。我们开发了针对公猪精子蛋白的单克隆抗体，可在体外诱导公猪精子凝集。