羰基咪唑法固定化pH梯度毛细管等电聚焦电泳用于蛋白质分离  

The Method of Immobilized pH Gradient of CIEF used in Protein Separation

作　　者：谢瑶[1] 耿利娜[1] 屈锋[1] 罗爱芹[1] 邓玉林[1] 

机构地区：[1]北京理工大学生命科学与技术学院,北京100081

出　　处：《化学通报》2008年第6期461-464,共4页Chemistry

基　　金：民用航天项目(D2220062902);国家自然科学基金项目(20705005)资助

摘　　要：本文以N,N′-羰基二咪唑作为连接臂,将两性电解质(CAs)与毛细管内壁进行偶联。在电场作用下,CAs在毛细管内按照等电点次序依次分开,再通过与羰基二咪唑的化学键合,形成固定化pH梯度。该方法操作简单,适用于两步法毛细管等电聚焦分离蛋白质等两性生物大分子。考察了对血红蛋白、牛血清白蛋白及胃蛋白酶三种蛋白质混合样品的分离,证明该方法可行。An immobilized pH gradient method had been developed for proteins separation by two-step capillary isoelectric focusing based on chemical bonding. The double functional reagent, N, N＇-carbonyldiimidazole was introduced in the capillary as a linkage in the immobilization reactions. Moreover, the commercial ampholytes （CAs） were distributed according to their pI values in the condition of electric field, and then connected with N, N＇-carbonyldiimidazole by chemical bonding to realize the immobilized pH gradient in capillary. A test sample containing a protein mixture of hemoglobin （Hb）, bovine serum albumin（BSA） and pepsin was separated by using this method.

关 键 词：固定化pH梯度 N N-羰基二咪唑 毛细管等电聚焦蛋白分离 

分 类 号：Q51-3[生物学—生物化学]