凝集素修饰的杂化亲和整体柱制备及糖蛋白质分离研究.docVIP

  凝集素修饰的杂化亲和整体柱制备及糖蛋 白质分离研究# 庞纪磊1,2,3，黄慧1,2,3，肖云1,2,3，林子俺1,2,3** 5 10 （1. 食品安全分析与检测教育部重点实验室，福州 350002； 2. 福建省食品安全分析与检测技术重点实验室，福州 350002； 3. 福州大学化学化工学院，福州 350002） 摘要：采用溶胶-凝胶法，以四乙氧基硅烷（TEOS）和 3-氨丙基三乙氧基硅烷（APTES）作 为反应前驱体, 以十六烷基三甲基溴化铵（CTAB）为超分子模板，在碱催化进行原位缩聚 反应, 再通过戊二醛偶联将植物血球凝集素修饰于硅胶基质表面，制备了新型的凝集素有机 -无机杂化亲和毛细管整体柱。优化了整体柱的制备条件，并对整体柱的形貌、孔径分布和 反应过程进行了表征。分别考察了前驱体比例、CTAB 含量对柱床结构和柱背压的影响。制 得的亲和整体柱对 N 型糖蛋白呈现出良好的选择性，可用于糖蛋白与非糖蛋白的分离。 关键词：液相色谱；杂化亲和整体柱；凝集素；糖蛋白质 15 中图分类号：O657 文献编识码：A  Preparation of lectin-based hybrid affinity monolithic column and its application for selective separation of  20 25 30 35 40 glycoprotein Pang Jilei1,2,3, Huang Hui1,2,3, Xiao Yun1,2,3, Lin Zian1,2,3 (1. Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, FuZhou 350002; 2. Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, FuZhou 350002; 3. College of Chemistry Chemical Engineering Fuzhou University, FuZhou 350002) Abstract: A lectin-based organic-inorganic hybrid monolithic column was prepared in a pretreated capillary by a sol-gel approach In this process, tetraethoxysilane (TEOS) and aminopropyltriethoxysilane (APTES) were used as the precursors, and then phytohaemagglutinin (PHA) was modified at the surface of the silica matrix by covalently binding glutaraldehyde. The prepared monolith was optimized and the morphology, permeability, and pore properties of the hybrid monolithic columns were characterized. The effect of salt concentration and methyl-α-D-mannopyranosideconcentrationwere on the elution of glycoproteins was examined. The result validated that the lectin-based hybrid affinity monolith had high selectivity towards N-glycoproteins. Keywords: Liquid chromatography; hybrid affinity monolithic column; lectin; glycoprotein 0 引言 凝集素（Lectin）是一类从动植物体内提取的糖蛋白或结合糖的蛋白，因其对含有糖链 结构的糖蛋白具有特异性亲和能力，长期以来被用于糖蛋白质的分离富集研究[1-4]。 目前，利用凝集素亲和原理分离富集糖蛋白的基质主要包括磁性纳米球[5]、琼脂糖[6-8]、 硅胶[9-10]、聚合物材料[11]和亲和膜[12]等。其中，琼脂糖亲和层析是利用琼脂糖固载的凝集素 装填柱子，通过色谱层析方法实现糖蛋白