人红细胞膜血型糖蛋白A的纯化及鉴定

The purification and identification of glycophorin A of human red blood cell membrane

目的：获得纯度高、抗原特异性好的Ｍ和Ｎ型血型糖蛋白Ａ（ＧＰＡ）。方法：将人红细胞膜血影，用ＳＤＳ溶解后用氯仿－甲醇提取，经ＳｅｐｈａｄｅｘＧ－１００凝胶过滤和ＷＧＡ－Ｓｅｐｈａｒｏｓｅ４Ｂ亲和层析。结果：获得了纯度高而均一的Ｍ，Ｎ型ＧＰＡ蛋白，ＳＤＳ－ＰＡＧＥ分析ＧＰＡ二聚体和单体相对的分子质量分别为７５×１０３，３８×１０３。结论：酶联免疫吸附分析（ＥＬＩＳＡ）和Ｗｅｓｔｅｒｎ印迹分析均表明，Ｍ和Ｎ型ＧＰＡ有很好的抗原性。

Objective: To prepare highly pure and good antigenspecific glycophorin A (GPA).Methods: GPA was purified from MM and NN blood group erythrocytes respectively by SDS solubilization,chloroformmethanol mixture extraction,Sephadex G100 filtration and WGASepharose4B affinity chromatography.Results: Highly pure and homogeneous M and NGPAs were obtained. Analysis showed that the molecular weight of GPA dimer and monomer were 75kD and 38kD respectively. Conclusion: Purified M, N GPA had good activity to AntiM and Anti-N McAbs respectively, but the antigenicity of NGPA was lower, suggesting that N GPA is more prone to change in its conformation than M GPA.