Brunner M, Wachtel S, Chin S
Center for Reproductive Biology, Collierville, TN 38017.
J Immunoassay. 1988;9(2):105-24. doi: 10.1080/15321818808057035.
Analysis of the male-specific H-Y antigen is difficult, in part, because of the tendency of "male-specific" antisera to bind cell surface components found in male and in female cells. Some insight as to the nature of that difficulty is provided by biochemical evaluation of H-Y antibody. In this study, purification of a monoclonal H-Y antibody with ammonium sulfate or Protein-A Sepharose, revealed the possibility of microheterogeneity. Despite evidence of multiple subtypes, Protein-A elution profiles suggested that the male-specific activity of the antibody resided in an IgG2a moiety. This was borne out by decreased activity after absorption of the IgG2a subtype with male cells, and by reaction of the monoclonal antibody with mouse subtype-specific antisera in an ELISA. Combined analysis using biological (absorption), biochemical (EF and PAGE) and immunological (ELISA) methods could find applicability in other complex systems.
对雄性特异性H-Y抗原的分析存在困难，部分原因是“雄性特异性”抗血清倾向于结合雄性和雌性细胞中都存在的细胞表面成分。对H-Y抗体的生化评估为了解该困难的本质提供了一些线索。在本研究中，用硫酸铵或蛋白A琼脂糖纯化单克隆H-Y抗体，揭示了微异质性的可能性。尽管有多种亚型的证据，但蛋白A洗脱曲线表明该抗体的雄性特异性活性存在于IgG2a部分。用雄性细胞吸收IgG2a亚型后活性降低，以及在酶联免疫吸附测定（ELISA）中该单克隆抗体与小鼠亚型特异性抗血清发生反应，都证实了这一点。综合使用生物学（吸收）、生化（电泳聚焦和聚丙烯酰胺凝胶电泳）和免疫学（ELISA）方法的分析可能适用于其他复杂系统。
