雷公藤内酯醇人工抗原的合成及多克隆抗体的制备  

Preparation of the triptolide-protein conjugate and the polyclonal antibody against triptolide

作　　者：唐波[1] 刘晓[1] 徐晓昱[1] 姚其正[2] 郑伟娟[1] 华子春[1] 

机构地区：[1]南京大学医药生物技术国家重点实验室,江苏南京210093 [2]中国药科大学药学院,江苏南京210009

出　　处：《东南大学学报（医学版）》2008年第6期409-414,共6页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(30425009;30730030);江苏省自然科学基金资助项目(BK2007715)

摘　　要：用碳二亚胺(EDC)法将14位羟基修饰的雷公藤内酯醇(TP)和不同的蛋白载体(阳离子化牛血清白蛋白、鸡卵清蛋白)偶联合成TP的人工免疫抗原和检测抗原,紫外光谱鉴定偶联效果,计算偶联率。利用免疫抗原免疫小鼠,制备小鼠多克隆抗体,用检测抗原分析血清抗体效价,利用抗原竞争ELISA分析抗体特异性,为进一步研究TP的分子作用机理以及制备TP的单克隆抗体奠定基础。The triptolide 14′-succinate was synthesized from the succinyl anhydride and triptolide, then the derivative was conjugated to the cBSA protein carrier to produce a suitable immunogen, using the EDC method. OVA was used to synthesize the analytical immunogen. The conjugation was proven to be successfully validated by UV spectral analysis. BALB/c mice were immunized by this conjugate and the polyclonal antibody against triptolide was prepared. The titer of the polyclonal antibody was measured and the specificity of the polyclonal antibody was verified. The preparation of polyclonal antibody against triptolide provides a useful tool for further preparation of monodonal antibody against triptolide and elucidation of molecular mechanism of triptolide function.

关 键 词：雷公藤内酯醇 14位羟基修饰 人工抗原 多克隆抗体 

分 类 号：Q503[生物学—生物化学]