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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.
Using bovine hemoglobin as the template protein, acrylamide as the functional monomer, N, N’-methylenebisacrylamide as the crosslinking monomer and ammonium persulfate as the initiator, the bovine hemoglobin molecule was prepared using the “sandwich” method The best experimental conditions were determined.The molar ratio of functional monomer to crosslinking agent was 40: 1, the eluent was 1% SDS-10% HAc, the elution rate of template protein was 47% The results showed that the adsorbed amount of template protein reached the maximum at 3.5 h and 2.9 μmol / L of recombinant protein, and the adsorption amount of template protein (0.22 nmol / cm2) was obviously (0.10 nmol / cm2) higher than that of non-imprinted membrane, and the values of imprinting factor for hemoglobin, myoglobin, lysozyme and bovine serum albumin were 2.5, 1.7, 1.3 and 1.2, respectively, Has a certain selectivity.