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目的 提取人及大鼠血清补体C5b-9复合物并制备抗体。方法 利用兔红细胞能直接活化人补体旁路途径的特性首先获得人或大鼠补体攻击破裂的兔红细胞膜 ,然后用脱氧胆酸钠溶解后经超速离心提取C5b-9复合物。结果 纯化的C5b -9复合物电镜负染观察有其独特的形态。免疫组化鉴定显示 ,抗C5、抗C9及抗C5b -9抗体染色阳性。另用鼠C5b -9复合物免疫家兔可制备相应的抗体。结论 补体旁路激活法提取C5b-9复合物快速简捷 ,结果可靠。
Objective To extract human and rat serum complement C5b-9 complex and prepare antibody. Methods Using the characteristics of rabbit erythrocytes that directly activated human complement pathway, we first obtained human or rat complement to attack the ruptured rabbit erythrocyte membrane and then the C5b-9 complex was extracted by ultracentrifugation after dissolving with sodium deoxycholate. Results The electron microscope observation of purified C5b-9 complex had its unique morphology. Immunohistochemical identification showed that anti-C5, anti-C9 and anti-C5b-9 antibodies were positive for staining. Another rat C5b -9 complex immunized rabbits can be prepared by the corresponding antibodies. Conclusion Complement bypass activation of C5b-9 complex is fast and simple and reliable.