“三明治”法制备牛血红蛋白分子印迹膜

"Sandwich-Prepare" of a Bovine Hemoglobin Molecularly Imprinted Polymer Membrane

以牛血红蛋白为模板蛋白、丙烯酰胺为功能单体、N,N′-亚甲基双丙烯酰胺为交联单体、过硫酸铵为引发剂,采用"三明治"法制备了牛血红蛋白分子印迹膜.确定了最佳的实验条件,功能单体与交联剂的摩尔配比为40∶1,洗脱剂为1%SDS-10%HAc,其对模板蛋白的洗脱率为47%.结果表明,所制备的蛋白质印迹膜在3.5 h和2.9μmol/L重结合蛋白浓度时,对模板蛋白的吸附量达到最大.该分子印迹膜对模板蛋白的吸附量(0.22 nmol/cm2)明显高于非印迹膜对模板蛋白的吸附量(0.10 nmol/cm2),对血红蛋白、肌红蛋白、溶菌酶和牛血清白蛋白的印迹因子值分别为2.5,1.7,1.3和1.2,说明它对模板蛋白具有一定选择性.

A hemoglobin molecularly imprinted polymer membrane（MIPM）was prepared on a glass chip by using water soluble acrylamide as a functional monomer and N,N′-methylenebisacrylamide as a crosslinker.The optimal ratio of functional monomer to crosslinker monomer to form a good membrane is 40∶1.1%SDS-10%HAc was the most effective washing solution which can remove 47% of the template.The absorption reached a maximal value at 3.5 h and at 2.9 μmol/L rebinding concentration.The MIPM showed higher absorb-capacity（0.22 nmol/cm2） to the template protein than that（0.10 nmol/cm2） of the non-imprinted polymer membrane（NIPM）.The selectivity experiment carried out in different protein solutions showed that MIPM absorbed more template protein than those that had not been imprinted.The imprinting factor for hemoglobin,myoglobin,lysozyme and BSA were 2.5,1.7,1.3 and 1.2 respectively.