摘要
目的：纯化制备兔源性抗人精子多克隆抗体标准品。方法：利用人精子抗原及其他组织蛋白等分别制备亲和层析柱，经正反相互补式亲和层析从兔抗血清中纯化IgG和IgA型抗人精子多克隆抗体，用SDS—PAGE、Western blot和ELISA等法检测其纯度、特异性和与精子抗原的线性反应。结果：所得抗体与牛血清白蛋白、正常人全血清、正常人组织细胞裂解液等在诊断试剂中常有的物质无可见交叉反应，在7．8120～0．4882ng·ml^-1或31．250～1．953ng·ml^-1浓度范围内与精予抗原的酶联吸附试验呈直线反应。结论：该纯化方案可用于抗精子抗体定量检测中抗体标准品的制备。
Objective To prepare purified rabbit anti-human sperm polyelonal antibody standard. Methods Three type of affinity chromatography column respectively made from human sperm antigen, normal serum and lysate of normal tissue cells were used to purify the anti-sperm polyclonal antibody（ IgG and IgA） from rabbit anti-human sperm serum. The resulting antibody was further applied to SDS-PAGE, western blot and enzyme linked immunoabsorbent assay （ELISA） for determining its purity, specificity and linear reaction with sperm antigen. Results The purified anti-sperm antibody shows no detectable cross reaction in western blot with BSA, human serum,lysate of human normal tissue cells which usually existed in immunodiagnostic reagents. At the concentration range of 0. 488 2 - 7. 812 0 ng ·ml^-1 （for IgG isotype） or 1. 953 - 31. 250 ng·ml^-1（for IgA isotype）, the purified antibody reacted in ELISA with sperm antigen in a linear manner. Conclusion The purification schedule is feasible for preparation of antibody standard used in antibody quantitative detection.
出处
《东南大学学报（医学版）》
CAS
2009年第6期500-504,共5页
Journal of Southeast University(Medical Science Edition)
关键词
抗精子抗体
定量酶联免疫吸附试验
亲和层析
anti-sperm antibody
quantitative enzyme-labeled immunosorbent assay
affinity chromatography
