Shigeta M, Watanabe T, Maruyama S, Koyama K, Isojima S
Clin Exp Immunol. 1980 Dec;42(3):458-62.
Rat spleen cells immunized to human azoospermic semen (a mixture of seminal plasma components) and mouse myeloma cells (P3/X63 Ag8U1; P3U1) (Marguilies et al., 1976) were successfully fused with polyethylene glycol (PEG 1500) and 19 of 89 fused cell cultures were found to produce sperm-immobilizing antibody. The cells that produced antibody indicating the highest sperm-immobilizing activity were distributed into wells for further recloning and 10 clones producing sperm-immobilizing antibody were established. The clone (1C4) producing the highest antibody titre was found to produce a large amount of IgG in culture supernatants and to contain a mixture of rat and mouse chromosomes. It was proved by immunodiffusion test that the monoclonal antibody was produced to the human seminal plasma antigen No. 7 which is common to human milk protein. Using this hybridoma which produced a large amount of monoclonal sperm-immobilizing antibody, a new method could be developed for purifying human seminal plasma antigen by immunoaffinity chromatography with bound antibody from the hybridoma.
用人类无精子症精液(精浆成分的混合物)免疫的大鼠脾细胞与小鼠骨髓瘤细胞(P3/X63 Ag8U1;P3U1)(Marguilies等人,1976年)成功地用聚乙二醇(PEG 1500)进行了融合,在89个融合细胞培养物中,有19个被发现产生精子制动抗体。将产生具有最高精子制动活性抗体的细胞分到孔中进行进一步的亚克隆,并建立了10个产生精子制动抗体的克隆。发现产生最高抗体滴度的克隆(1C4)在培养上清液中产生大量IgG,并且含有大鼠和小鼠染色体的混合物。免疫扩散试验证明,该单克隆抗体是针对人乳蛋白中常见的人类精浆抗原7产生的。利用这种产生大量单克隆精子制动抗体的杂交瘤,可以开发一种新方法,即通过用来自杂交瘤的结合抗体进行免疫亲和层析来纯化人类精浆抗原。