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目的 :检测OPG与TNF家族蛋白的结合及探索它们相互作用机制。方法 :免疫沉淀分析加Western印迹 ,酶联免疫吸附测定方法 ,偶联剂交叉链接 ,表面胞质团共振。结果 :OPG与RANKL ,TRAIL和TNFα用不同方法检测均有结合。OPG与TNFα的结合由Biacore进一步证实 ,在 pH为 7.4 ,温度在 2 0℃时 ,其离解系数为7.0nmol/L ,同OPG TRAIL亲和力。偶联剂交叉链接显示OPG TNFα结合的比例为 1∶1。结论 :TNFα能与OPG结合 ,物理条件影响其结合。
OBJECTIVE: To detect the binding of OPG to TNF family proteins and to explore their mechanisms of interaction. Methods: Immunoprecipitation analysis plus Western blotting, enzyme linked immunosorbent assay, cross-linking of coupling agents, surface plasmon resonance. Results: OPG and RANKL, TRAIL and TNFα were detected by different methods. The binding of OPG to TNFα was further confirmed by Biacore, with a dissociation coefficient of 7.0 nmol / L at pH 7.4 and a temperature of 20 ° C, which shows affinity with OPG TRAIL. Coupling agent cross-linking showed a 1: 1 ratio of OPG TNF [alpha] binding. Conclusion: TNFα can bind to OPG and its physical conditions affect its binding.