Rosenberry T L, Chen J F, Lee M M, Moulton T A, Onigman P
J Biochem Biophys Methods. 1981 Jan;4(1):39-48. doi: 10.1016/0165-022x(81)90004-x.
A molecular filtration procedure for preparing large quantities of human erythrocyte ghost membranes is presented. Hemolysate ghost membranes are rapidly cycled in the retentate channel of the filtration apparatus, while hemoglobin is removed as it passes through Pellicon filters into the filtrate channel. Several-liter quantities of washed packed erythrocytes can be processed in a few hours with this system, and the filtration procedure does not appear to alter intact erythrocyte or ghost membranes. Intact erythrocytes in isotonic solution can be circulated through the retentate channel for 16 h with only 3% hemolysis and with preservation of their original morphology in scanning electron microscopy. Ghost membranes isolated by the procedure are virtually identical in morphology, polypeptide composition and acetylcholinesterase content to membranes isolated by conventional centrifugation techniques.
本文介绍了一种用于大量制备人红细胞血影膜的分子过滤方法。溶血后的血影膜在过滤装置的截留液通道中快速循环,而血红蛋白在通过百得滤器进入滤液通道时被去除。使用该系统,几升洗涤后的压积红细胞可在数小时内处理完毕,且过滤过程似乎不会改变完整红细胞或血影膜。等渗溶液中的完整红细胞可在截留液通道中循环16小时,溶血率仅为3%,且在扫描电子显微镜下能保持其原始形态。通过该方法分离得到的血影膜在形态、多肽组成和乙酰胆碱酯酶含量方面与通过传统离心技术分离得到的膜几乎相同。