Skuse D R, Müller W, Brooks D E
Department of Chemistry, University of British Columbia, Vancouver, Canada.
Anal Biochem. 1988 Nov 1;174(2):628-35. doi: 10.1016/0003-2697(88)90066-8.
Cell separation using aqueous polymeric two-phase systems is well established. For separations of cells having similar partition coefficients a multistep countercurrent distribution procedure has to be used. However, its operation is limited by time and apparatus constraints. As an alternative strategy we have developed a chromatographic technique in which the dextran-rich phase of a dextran/polyethylene glycol (PEG) phase system is immobilized onto derivatized agarose beads. The PEG-rich phase is used as the eluent. Inclusion of PEG-fatty acid affinity ligand gradients into the eluent produces separations of mammalian erythrocytes based on the differential interaction between the fatty acid and the erythrocyte membranes. A model separation of dog and human erythrocytes has been carried out.
使用水性聚合物双相系统进行细胞分离已得到充分确立。对于具有相似分配系数的细胞分离，必须使用多步逆流分配程序。然而，其操作受到时间和设备限制。作为一种替代策略，我们开发了一种色谱技术，其中葡聚糖/聚乙二醇（PEG）相系统中富含葡聚糖的相被固定在衍生化的琼脂糖珠上。富含PEG的相用作洗脱液。在洗脱液中加入PEG-脂肪酸亲和配体梯度，可基于脂肪酸与红细胞膜之间的差异相互作用对哺乳动物红细胞进行分离。已对犬和人红细胞进行了模型分离。
