“Grafting from”法制备高载量大孔弱阳离子交换层析介质  

Preparation of Macroporous Weak Cation Exchange Chromatography Medium with High Capacity through“Grafting from”Method

作　　者：郭旺 姜泽平 马磊 乔娟 莫文清 靳海波[1] 何广湘[1] 黄永东[2] 张荣月[1] GUO Wang;JIANG Zeping;MA Lei;QIAO Juan;MO Wenqing;JIN Haibo;HE Guangxiang;HUANG Yongdong;ZHANG Rongyue(Beijing Key Laboratory of Clean Fuel and Efficient Catalytic Emission Reduction Technology,College of New Materials and Chemical Engineering,Beijing University 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：《高等学校化学学报》2024年第3期64-72,共9页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:22074148);国家重点研发计划项目(批准号:2021YFC2103401)资助.

摘　　要：针对大孔聚合物层析介质孔径大、比表面积低而导致的蛋白结合容量低的问题,采用“graftingfrom”策略,以大孔聚丙烯酸酯微球为基质,通过氧化还原引发甲基丙烯酸在微球表面接枝聚合,制备了高载量弱阳离子交换层析介质.研究了单体浓度、过硫酸钾浓度及反应温度等因素对蛋白结合容量的影响.所得介质的蛋白静态结合容量和动态结合容量分别达到252.21和157.25mg/mL;同时发现具有一定离子交换容量的该类介质能够在0.2 mol/L NaCl缓冲液中保持100 mg/mL的蛋白结合容量.将该层析介质用于鸡卵清中溶菌酶的纯化,获得了较高的纯化效率.Macroporous polymer chromatography medium has a low protein binding capacity due to its large pore size and low surface area.To solve this problem,the weak cation exchange chromatography medium with high capacity was prepared through the grafting polymerization of methacrylic acid on the macroporous polyacrylate microspheres,which was initiated by redox reaction.The reaction factors were evaluated and the general rules between these factors and proteins binding capacity were obtained,including monomer concentration,potassium persulfate concentration and reaction temperature.The static binding capacity and dynamic binding capacity of the obtained medium reached 252.21 and 157.25 mg/mL,respectively.It was also found that the medium with a certain ion exchange capacity could maintain 100 mg/mL protein binding capacity in 0.2 mol/L NaCl buffer.The chromatographic medium was used for the purification of lysozyme in egg white,and high purification efficiency was obtained.

关 键 词：弱阳离子交换层析介质 大孔聚合物 蛋白结合容量 Grafting from法 分离纯化 

分 类 号：O658.1[理学—分析化学] O632.1[理学—化学]