Dimitrova D, Kehayov I, Kyurkchiev S
Department of Biology, Medical Faculty, Sofia, Bulgaria.
Andrologia. 1993 Sep-Oct;25(5):271-7. doi: 10.1111/j.1439-0272.1993.tb02724.x.
A procedure was designed for purification of a human seminal plasma-specific antigen (HSP-antigen) identified by a monoclonal antibody produced in this laboratory (mAb 4E6). Pooled human seminal plasma was fractionated by consecutively applied methods: affinity chromatography on Lentil lectin sepharose, gel chromatography on Ultrogel AcA 34 and immunoaffinity on mAb 4E6 coupled CNBr-Sepharose 4B. The antigen-containing fraction obtained after this procedure was proved to be homogeneous when analysed by electrophoresis in polyacrylamide gel. After the consecutive purification procedures the degree of purification was 128 times as compared to the starting material. Electrophoretic analysis of the purified 4E6 antigen under reducing conditions showed that it consisted of 3 polypeptide subunits with molecular weight 70 kDa, 64 kDa and 60 kDa respectively. On the basis of the data obtained from competitive ELISA testing of sera from infertile patients it has been suggested that the identified antigen may be involved in pathogenesis of immunologic infertility.
设计了一种程序，用于纯化由本实验室产生的单克隆抗体（mAb 4E6）鉴定的人精浆特异性抗原（HSP抗原）。通过连续应用的方法对合并的人精浆进行分级分离：在扁豆凝集素琼脂糖上进行亲和层析、在Ultrogel AcA 34上进行凝胶层析以及在偶联有mAb 4E6的CNBr-琼脂糖4B上进行免疫亲和层析。当通过聚丙烯酰胺凝胶电泳分析时，该程序后获得的含抗原部分被证明是均一的。经过连续纯化程序后，纯化程度与起始材料相比为128倍。在还原条件下对纯化的4E6抗原进行电泳分析表明，它由分别具有70 kDa、64 kDa和60 kDa分子量的3个多肽亚基组成。基于对不育患者血清进行竞争性ELISA检测获得的数据，有人提出所鉴定的抗原可能参与免疫性不育的发病机制。
