中文摘要：

目的：运用亲和层析方法初步探索栀子苷的特异结合蛋白。方法：采用环氧氯丙烷（Epichlorohydrin,ECH）作为活化试剂对琼脂糖凝胶进行环氧基修饰,通过偶联栀子苷与环氧活化的琼脂糖凝胶（epoxy activated Sepharose CL-6B,EAS6B）,制备以栀子苷为配基的亲和介质,以高效液相色谱法验证其偶联。利用栀子苷-EAS6B亲和介质,从小鼠脑组织总蛋白中筛选特异结合蛋白。结果：优化偶联条件后,得到最佳的活化反应条件：15%ECH、0.8 mol/L Na OH于37℃反应3 h,环氧基密度达到122μmol/m L,琼脂糖凝胶亲和介质的偶联量为19.53%,偶联率为17.08μmol/m L。从小鼠脑组织总蛋白中筛选得到3个栀子苷特异结合蛋白的条带,经质谱鉴定,发现主要是由热休克蛋白、脑酸溶性蛋白和谷氨酰胺合成酶等组成。结论：确立了琼脂糖凝胶环氧活化的最佳条件,制备了栀子苷-EAS6B亲和介质并应用其筛选出栀子苷的结合蛋白。

英文摘要：

Objective： To preliminary identify the specific binding protein of geniposide using affinity chromatography. Methods：ECH, the abbreviation for Epichlorohydrin, was adopted as activating reagent for surface modification of agarose gel（epoxy activated Sepharose CL-6B, EAS6B）. Geniposide was coupled with EAS6 B by hydroxyl-epoxy interaction, and high performance liquid chromatography was put into use to verify the efficiency of coupling, and then geniposide-EAS6 B was applied to screening the binding protein of geniposide in mouse brain. Results： The optimal condition of epoxy activated Sepharose CL-6B was as followed： 15 % ECH, 0.8 mol/L Na OH was reacted at 37 ℃ for 3 h. After optimized the conditions, the epoxy group density was up to 122 μmol/m L, the conjugation ratio and amount were 19.53 % and 17.08 μmol/m L, respectively. Three bands on SDS-PAGE were obtained from total protein in mouse brain using affinity chromatography. Conclusions： We established the optimal condition of epoxy activated Sepharose CL-6B, and prepared the geniposide-EAS6 B affinity resin that was applied to screening the binding proteins of geniposide.