烯丙基活化制备大孔强阳离子交换介质及其在蛋白分离中的应用  

Preparation of Macroporous Strong Cation Exchange Chromatography Medium by Allyl Activation and Its Application in Protein Separation

作　　者：薛彦晓 侯恒扬 公丕胜 靳海波[1] 赵岚[2] 何广湘[1] 张荣月[1] XUE Yan-Xiao;HOU Heng-Yang;GONG Pi-Sheng;JIN Hai-Bo;ZHAO Lan;HE Guang-Xiang;ZHANG Rong-Yue(Beijing Key Laboratory of Fuels Cleaning and Advanced Catalytic Emission Reduction Technology,College of New Materials and Chemical Engineering,Beijing Institute of Petrochemical Technology,Beijing 102617,China;State Key Laboratory of Biochemical Engineering,Institute of Process Engineering,Chinese Academy of Sciences,Beijing 100190,China)

机构地区：[1]北京石油化工学院新材料与化工学院,燃料清洁化及高效催化减排技术北京市重点实验室,北京102617 [2]中国科学院过程工程研究所,生化工程国家重点实验室,北京100190

出　　处：《分析化学》2024年第12期1896-1903,共8页Chinese Journal of Analytical Chemistry

基　　金：北京市委组织部青年拔尖人才项目(No.ZZB2019001);中国科学院化学研究所活体分析化学中国科学院重点实验室开放课题项目(No.ACL201903);北京市自然科学基金项目(No.L192017)资助。

摘　　要：强阳离子交换介质广泛应用于蛋白的分离与纯化。本研究以甲基丙烯酸缩水甘油酯和乙二醇二甲基丙烯酸酯共聚大孔微球为基质,经烯丙基活化偶联小分子配体,制备了聚合物基质的强阳离子交换介质。系统考察了各反应因素对配基密度的影响,制备得到配基密度在0.126~0.370 mol/L范围内的介质,并对其吸附性能进行了评价。以溶菌酶为模型蛋白,测得其静态载量范围为37.0~81.0 g/L,动态载量为37.7 g/L,溶菌酶的最高回收率达到98.97%。此介质具有较低的非特性吸附,并且对溶菌酶有良好的分离效果,在快速蛋白纯化方面有良好的应用前景。Strong cation exchange mediums have been widely applied in protein purification.In this work,the strong cation exchange chromatography medium was prepared by coupling small molecular ligands through allyl activation commonly used in agarose-based media.The matrix based on the macroporous microspheres was a copolymer of glycidyl methacrylate and ethylene glycol dimethacrylate.The effects of reaction factors on the ligand density were detailedly investigated.The ligand density in the prepared medium was controlled in the range of 0.126-0.370 mol/L(wet gel).Its adsorption performance was evaluated with lysozyme as a model protein.The static binding capacity was in the range of 37.0-81.0 g/L.The maxium dynamic binding capacity reached 37.7 g/L and the maximum lysozyme recovery was 98.97%.The strong cation exchange medium obtained here showed low nonspecific adsorption and high separation efficiency on lysozyme.This strong cation exchange medium showed a great potential in rapid protein purification.

关 键 词：强阳离子交换层析介质 烯丙基活化 配基密度 吸附性能 分离纯化 

分 类 号：TQ936[化学工程]