白眉蝮蛇蛇毒纤溶酶的聚乙二醇修饰  

Pegylation of a Fibrinolytic Enzyme from the Venom of Agkistrodon halys ussuriensis

作　　者：公伟[1] 朱裕辉[1] 薛春佳[1] 冯军[1] 赵文杰[1] 

机构地区：[1]上海医药工业研究院,上海200040

出　　处：《中国医药工业杂志》2007年第10期696-700,共5页Chinese Journal of Pharmaceuticals

摘　　要：为提高纤溶酶的临床适用性,采用4种针对氨基修饰的mPEG试剂—mPEG-SPA 5K、mPEG-SMB 5K、mPEG-ButyrALD 5K和mPEG2-NHS 40K修饰蛇毒纤溶酶,以获得高比例的单修饰产物和较高的酶活性保留率。优化修饰反应条件,用Superdex 75凝胶过滤色谱法分离纯化修饰产物。最终确定mPEG-SPA 5K为最适宜的修饰剂。产物经SDS-PAGE和MALDI-TOF-MS法检测为单修饰,修饰产物酶活性收率为67.3%,修饰产物的热稳定性提高,免疫原性显著下降。Four activated mPEG reagents （mPEG-SPA 5K, mPEG-SMB 5K, mPEG-ButyrALD 5K and mPEG2-NHS 40K） were selected to modify the amino groups in fibrinolytic enzyme （FIE） molecule. The pegylated conditions were optimized to obtain high proportion of mono-pegylated FIE and the relatively higher enzyme activity. mPEG-SPA 5K was finally confirmed as the best modifier for FIE. The pegylated FIE was separated by gel filtration chromatography and was confirmed to be mono-pegylated by SDS-PAGE and MALDI-TOF-MS. Its enzyme activity yield was 67.3 %. The mono-pegylated FIE showed higher thermo stability and greatly lower immunogencity.

关 键 词：纤溶酶 聚乙二醇修饰 纯化 

分 类 号：Q814[生物学—生物工程]