Lundwall A, Eggertsen G
J Immunol Methods. 1985 Jul 16;81(1):147-60. doi: 10.1016/0022-1759(85)90131-0.
An improved method for simultaneous purification of complement factors C3, C5 and H from human plasma has been developed. Using an initial batch separation technique with QAE-Sephadex, followed by chromatography on SP-Sephadex and gel filtration in Sephadex G-200, 600 mg of highly pure C3 can be prepared from 1600 ml of plasma. Simultaneously about 70 mg of highly pure factor H and 30 mg of C5 are obtained by chromatography of post SP-Sephadex material on DEAE-Sephacel. A small amount of C3 in the C5 pool is removed by anti-C3-Sepharose. By maleylation or citraconylation of reduced and alkylated C3, the constitutive polypeptide chains are modified in a way that made them separable by ion exchange chromatography.
已开发出一种从人血浆中同时纯化补体因子C3、C5和H的改进方法。采用QAE-葡聚糖凝胶的初始批量分离技术，随后进行SP-葡聚糖凝胶色谱和Sephadex G-200凝胶过滤，可从1600毫升血浆中制备出600毫克高纯度的C3。同时，通过将SP-葡聚糖凝胶柱后的物质在DEAE-琼脂糖凝胶上进行色谱分离，可获得约70毫克高纯度的因子H和30毫克C5。通过抗C3-琼脂糖凝胶去除C5组分中的少量C3。通过对还原和烷基化的C3进行马来酰化或柠康酰化，组成性多肽链以一种可通过离子交换色谱分离的方式被修饰。
