Elce J S, Graham E J, Zboril G, Leyton L, Perez E, Croxatto H B, de Ioannes A
Biochem Cell Biol. 1986 Dec;64(12):1242-8. doi: 10.1139/o86-163.
Monoclonal antibodies to human acrosin were required for studies of immunological interference with fertilization. Since human acrosin was not available in adequate amounts, monoclonal antibodies have been raised in mice against purified bovine acrosin and screened for cross-reaction with human sperm cells. Two of these antibodies are described, B4F6 and C2E5. Data from enzyme-linked immunosorbent assays, immunoblots, immunoprecipitation, and indirect immunofluorescence on sperm cells indicate that B4F6 binds only to bovine acrosin, and that C2E5 binds both to bovine and to human acrosin at a conformationally determined epitope. The antibodies do not inhibit the hydrolysis of benzoylarginine ethyl ester by acrosin, but C2E5 did inhibit the dissolution of the hamster zona pellucida by purified human acrosin. The antibodies have also been used for affinity purification of acrosin and proacrosin.
为了研究免疫对受精的干扰,需要针对人精子顶体蛋白酶的单克隆抗体。由于无法获得足够量的人精子顶体蛋白酶,因此在小鼠中制备了针对纯化牛精子顶体蛋白酶的单克隆抗体,并筛选其与人精子细胞的交叉反应。本文描述了其中两种抗体,即B4F6和C2E5。来自酶联免疫吸附测定、免疫印迹、免疫沉淀以及精子细胞间接免疫荧光的数据表明,B4F6仅与牛精子顶体蛋白酶结合,而C2E5在一个由构象决定的表位上既能与牛精子顶体蛋白酶结合,也能与人精子顶体蛋白酶结合。这些抗体不抑制精子顶体蛋白酶对苯甲酰精氨酸乙酯的水解,但C2E5确实抑制了纯化的人精子顶体蛋白酶对仓鼠透明带的溶解。这些抗体还被用于精子顶体蛋白酶和前精子顶体蛋白酶的亲和纯化。