溶菌酶分子印迹微球的制备与评价

Preparation and Evaluation of Lysozyme Molecularly Imprinted Microspheres

以丙烯酰胺为功能单体、N,N'-亚甲基双丙烯酰胺为交联剂、溶菌酶为模板蛋白,用反相悬浮聚合法制备了溶菌酶分子印迹微球。所得微球外观圆整,平均粒径为34.6μm,ζ电位为-33.5 mV。考察了溶菌酶印迹微球在水和生理盐水两种介质中对溶菌酶、核糖核酸酶A或牛血清白蛋白的吸附量,以及微球在生理盐水中对两种蛋白质混合溶液中溶菌酶的特异性吸附能力。结果表明,微球对溶菌酶的吸附在40 min内达到平衡。无论在单一蛋白质还是有其它蛋白质干扰的竞争环境中,印迹微球对模板蛋白溶菌酶都表现出更强的吸附能力。在生生理盐水介质中,由于降低了非特异性吸附效应,印迹微球对模板蛋白的选择性吸附更显著。

An inverse-phase suspension polymerization method was applied to prepare lysozyme （template protein） molecularly imprinted microspheres （Lyz-MIM） with acrylamide as fimctional monomer and N,N＇-methylene bisacrylamide as cross-linker. The results demonstrated that Lyz-MIM had a spherical morphology with an average particle size of 34.6μm and ζ potential of -33.5 mV. The adsorption capacities of Lyz-MIM toward lysozyme, RNase A or BSA in water and physiological saline as well as its selectivity toward template protein in physiological saline were investigated. The adsorption kinetics study indicated that the adsorption equilibrium of Lyz-MIM was within 40 min. Whether in single or binary protein competitive rebinding test, Lyz-MIM showed a preferential rebinding toward Lyz, which became more significant when measured in physiological saline, probably due to the reduction of non-specific adsorption.