Le Bec A, Waitz R
Rev Fr Transfus Immunohematol. 1976 Sep;19(3):395-413.
Human erythrocyte membranes were prepared by hypotonic hemolysis and extracted with water pH 7, EDTA and mercapto-ethanol solutions pH 8. The complex residue which contains lipids, proteins and the A, B, H antigens was dissolved in SDS and analysed by preparative acrylamide gel electrophoresis in the presence of SDS. Each fraction was assayed for A, B or H blood-group activity by hemagglutination inhibition tests. Two experiments were done for each blood-group A, B, O. One with entire ghosts, another after extraction of the ghosts residue with a mixture of chloroform-methanol. The comparison of results showed that the distribution of proteins and blood-group antigen activity among the different fractions is not modified and that, in the two experiments, the elution of H substance is delayed by comparison with elution of A and B substances. The blood-group A, B and H antigens are known to be glycosphingolipids and are not entirely eliminated by treatment of the ghosts with cloroform and methanol. It has recently been reported [7] that these antigens are complex glycosphingolipids with hydrophilic character and remain in the aqueous phase after extraction of the erythrocyte membrane with organic solvents.
人红细胞膜通过低渗溶血法制备，并用pH 7的水、EDTA和pH 8的巯基乙醇溶液进行提取。含有脂质、蛋白质以及A、B、H抗原的复合残留物溶解于SDS中，并在SDS存在的情况下通过制备性丙烯酰胺凝胶电泳进行分析。通过血凝抑制试验对每个组分进行A、B或H血型活性检测。对A、B、O每种血型进行了两个实验。一个实验使用完整的血影，另一个实验是在用氯仿 - 甲醇混合物提取血影残留物之后进行。结果比较表明，不同组分之间蛋白质和血型抗原活性的分布没有改变，并且在这两个实验中，与A和B物质的洗脱相比，H物质的洗脱延迟。已知A、B和H血型抗原是糖鞘脂，用氯仿和甲醇处理血影并不能将其完全去除。最近有报道[7]称，这些抗原是具有亲水性的复合糖鞘脂，在用有机溶剂提取红细胞膜后仍保留在水相中。
