强阳离子交换树脂在蛋白质分离纯化中的应用  

Strong cation exchange stationary phase on monosized hydrophilic non-porous resins and their application for fast separation of proteins

作　　者：卜春苗[1] 朱金霞[1] 龚波林[1] 

机构地区：[1]宁夏大学能源化工重点实验室,宁夏银川750021

出　　处：《宁夏工程技术》2006年第1期60-64,共5页Ningxia Engineering Technology

基　　金：国家科技部前期研究专项基金资助项目(2004CC05000);国家自然科学基金资助项目(20465001);教育部新世纪优秀人才支持计划(NCET-04-0980)基金资助项目

摘　　要：为了进一步考察流动相中盐种类、体积流量及有机溶剂等对蛋白质保留的影响,采用新的化学方法将3.0μm无孔单分散亲水性交联聚甲基丙烯酸环氧丙酯树脂改性为一种新型的强阳离子交换色谱填料;实验分析了该填料对标准蛋白的分离性能,有机溶剂、流动相盐种类和体积流量等对蛋白质保留的影响.实验结果表明:当体积流量为3mL/min时,在1.5min线性梯度时间内可快速分离四种标准蛋白质,且蛋白质的保留符合阳离子交换色谱规律.该填料的动力学吸附容量为7.13mg/mL,将其应用于快速纯化鸡蛋清中的溶菌酶和猪心中细胞色素-C,可取得较好的效果.A strong cation-exchange（SCX） stationary phase had been synthesized by a new chemical modification of the 3.0 μm non-porous monodisperse poly（glyciddylmethacrylate-co-ethylenedimethaerylate） resin as medium. The stationary phase was evaluated in detail to determine its ion-exchange properties, hydrophilieity, and the effects of pH, flow rate and organic solvent on the separation and retention of proteins. It was found that retention times of proteins all followed cation ion exchange chromatographic （IEC） retention mechanism. Four proteins could be separated in 1.5 minutes by flow rates at 3 mL/min. The SCX resin was used for the fast purification of lysozyme from egg white and eytoehrome-C from pig heart by only one step. The satisfactory results have been obtained.

关 键 词：无孔单分散亲水性交联聚甲基丙烯酸环氧丙酯树脂 强阳离子交换色谱 蛋白质分离 

分 类 号：TQ323[化学工程—合成树脂塑料工业] Q503[生物学—生物化学]