Johnson A J, MacDonald V E, Semar M, Fields J E, Schuck J C, Brind J
J Lab Clin Med. 1978 Aug;92(2):194-210.
The solid-phase ethylene/maleic anhydride PE's E-5 and E-100 were used to developed a flexible integrated system of batch methods for the fractionation of albumin and gamma globulin as well as the clinically important coagulation factors. Each method for the isolation and purification of the fractions or a modification of it could also be used independently or could be added onto another method or methods. Each fraction was prepared in three steps: adsorption on solid-phase PE's elution from the PE's, and concentration by ultrafiltration or by use of PEG or both. Since the methods required no organic solvent but relied primarily on electrostatic bonding, they were essentially nondenaturing. Solid-liquid phase separation was easily accomplished by centrifugation or filtration. The yield and purity of each fraction was as follows: (1) coagulation factors II, VII, IX, and X concentrates 63% and 200-fold purified; (2) AHF 43% and 52-fold; (3) vWF 43% and 71-fold; (4) PPF, as albumin, 93% yield and 92% pure; (5) albumin 92% yield and 98.5% pure; and (6) gamma globulin 92% yield and 97.5 pure. During the preparation of gamma globulin the E-100 also tended to adsorb and remove the HBSAg and infectivity, when present.
采用固相乙烯/马来酸酐聚乙烯(PE)的E - 5和E - 100开发了一种灵活的分批法集成系统,用于分离白蛋白、γ球蛋白以及临床上重要的凝血因子。每种分离和纯化各组分的方法或其改进方法也可单独使用,或添加到其他一种或多种方法上。每个组分的制备分三步进行:吸附于固相PE上、从PE上洗脱以及通过超滤或使用聚乙二醇(PEG)或两者结合进行浓缩。由于这些方法无需有机溶剂,主要依靠静电结合,所以基本上是非变性的。固 - 液分离通过离心或过滤很容易实现。各组分的产率和纯度如下:(1)凝血因子II、VII、IX和X浓缩物,产率63%,纯化200倍;(2)抗血友病因子(AHF),产率43%,纯化52倍;(3)血管性血友病因子(vWF),产率43%,纯化71倍;(4)作为白蛋白的前蛋白原片段(PPF),产率93%,纯度92%;(5)白蛋白,产率92%,纯度98.5%;(6)γ球蛋白,产率92%,纯度97.5%。在制备γ球蛋白的过程中,如果存在乙肝表面抗原(HBSAg)及其传染性,E - 100还倾向于吸附并去除它们。