Qin Lei, He Xi-Wen, Zhang Wei, Li Wen-You, Zhang Yu-Kui
Department of Chemistry, Nankai University, Tianjin 300071, China.
J Chromatogr A. 2009 Jan 30;1216(5):807-14. doi: 10.1016/j.chroma.2008.12.007. Epub 2008 Dec 7.
A new and facile fabricating method for lysozyme molecularly imprinted polymer beads (lysozyme-MIP beads) in aqueous media was presented. Mesoporous chloromethylated polystyrene beads (MCP beads) containing dithiocarbamate iniferter (initiator transfer agent terminator) were used as supports for the grafting of lysozyme imprinted copolymers with acrylamide and N,N'-methylenebisacrylamide through surface initiated living-radical polymerization (SIP). After the polymerization, a layer of lysozyme-MIP was formed on the MCP beads. The SIP allowed an efficient control of the grafting process and suppressed solution propagation. Therefore, the obtained lysozyme-MIP beads had a large quantity of well-distributed pores on the surface without any visible gel formation in solution and were more advantageous comparing with traditional MIPs which were prepared by traditionally initiated radical polymerization. The obtained composites were characterized by Fourier transform infrared spectroscopy, elemental analysis, nitrogen sorption analysis and scanning electron microscopy. Chromatographic behaviors of the column packed with lysozyme-MIP beads exhibited ability in separating lysozyme from competitive protein (bovine hemoglobin, bovine serum albumin, ovalbumin or cytochrome c) in aqueous mobile phase.
本文提出了一种在水介质中制备溶菌酶分子印迹聚合物微球(溶菌酶-MIP微球)的新型简便方法。以含有二硫代氨基甲酸酯引发转移终止剂(iniferter)的介孔氯甲基化聚苯乙烯微球(MCP微球)为载体,通过表面引发活性自由基聚合(SIP)接枝溶菌酶印迹共聚物与丙烯酰胺和N,N'-亚甲基双丙烯酰胺。聚合反应后,在MCP微球表面形成了一层溶菌酶-MIP。SIP能够有效控制接枝过程并抑制溶液中的链增长。因此,所制备的溶菌酶-MIP微球表面具有大量分布均匀的孔,溶液中无可见凝胶形成,与传统自由基聚合制备的传统MIP相比更具优势。通过傅里叶变换红外光谱、元素分析、氮吸附分析和扫描电子显微镜对所得复合材料进行了表征。填充溶菌酶-MIP微球的色谱柱在水相流动相中具有从竞争性蛋白质(牛血红蛋白、牛血清白蛋白、卵清蛋白或细胞色素c)中分离溶菌酶的能力。