Bhakdi S, Roth M
J Immunol. 1981 Aug;127(2):576-80.
A rapid, simple method for isolating SC5b-9 generated in human serum, and SC5b-8 generated in C9-depleted serum is described. The procedure leads to approximately 30% recovery of either complex and consists of polyethyleneglycol precipitation followed by 1 DEAE ion exchange chromatography and a sucrose density gradient ultracentrifugation. The SC5b-8 complex is a water-soluble macromolecular of alpha-electrophoretic mobility. Apart from the absence of C9, it exhibits an identical SDS-gel electrophoresis polypeptide pattern as SC5b-9. A sedimentation coefficient of 19 to 20S and an effective molecular radius of approximately 10 nm were determined by sucrose density gradient centrifugation and gel filtration, respectively. From these values, a m.w. of 800 to 850,000 is tentatively assigned to the SC5b-8 complex. The S-protein was not dissociable from either SC5b-8 or SC5b-9 through treatment with desoxycholate and no alterations in the hydrodynamic properties of either complex were discernible after action of this detergent.
本文描述了一种快速、简单的方法，用于分离人血清中产生的SC5b-9以及C9缺失血清中产生的SC5b-8。该方法可使两种复合物的回收率约为30%，包括聚乙二醇沉淀，随后进行1次DEAE离子交换色谱和蔗糖密度梯度超速离心。SC5b-8复合物是一种具有α-电泳迁移率的水溶性大分子。除了不含C9外，它与SC5b-9具有相同的SDS-凝胶电泳多肽图谱。通过蔗糖密度梯度离心和凝胶过滤分别测定其沉降系数为19至20S，有效分子半径约为10nm。根据这些值，初步确定SC5b-8复合物的分子量为800至850,000。通过脱氧胆酸盐处理，S蛋白与SC5b-8或SC5b-9均不能解离，并且在该去污剂作用后，两种复合物的流体动力学性质均未出现明显变化。
