固载蛋白A交联聚甲基丙烯酸缩水甘油酯连续床亲合色谱柱的制备及性能考察

摘　要：采用沉淀聚合法“原位”(in-situ)聚合合成了交联聚甲基丙烯酸缩水甘油酯连续床色谱柱，对其进行化学改性后，分别得到含有11个碳原子间隔臂以及不含间隔臂键合了蛋白A的高效亲合色谱柱。考察了这两种色谱介质的性能，结果表明含有间隔臂的介质有一定的疏水性；对传统制备蛋白配基的亲合色谱介质的合成路线进行了改进；采用不含间隔臂的亲合柱测定了人血浆中人免疫球蛋白G(HIgG)的含量，所测得的定量标准曲线线性相关系数达到0.999；考察了流速对所合成的连续床色谱柱柱压的影响，当流速高达9.0mL/min时，柱压也仅为6.5MPa；采用pH梯度对HIgG的不同亚基进行了分离。

关键词：亲合色谱法，聚合物，连续床色谱柱，蛋白A，人免疫球蛋白G

分类号：O65　文献标识码：A

Preparation and Characterization of Protein A Immobilized on Molded Monolithic Rod of Macroporous Poly(glycidyl methacrylate-co-ethylene dimethacrylate)for Affinity Chromatography

Luo Quanzhou（National Chromatographic Research & Analysis Centre,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116011）　
Zou Hanfa（National Chromatographic Research & Analysis Centre,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116011）　
Wang Hailin（National Chromatographic Research & Analysis Centre,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116011）　
Mao Xiqin（National Chromatographic Research & Analysis Centre,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116011）　
Kong Liang（National Chromatographic Research & Analysis Centre,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116011）　
Ni Jianyi（National Chromatographic Research & Analysis Centre,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116011）　

Abstract：Reactive continuous rods of macroporous poly(glycidyl methacrylate-co-ethylene dimethacrylate) have been prepared by “in-situ” copolymerization of the monomers in the presence of porogenic dilute,and protein A was immobilized on the continuous rods directly or through a 11 carbon atom space arm with molded synthetic methods.The properties of these two rods columns were characterized and the results showed that the medium with space arm had some extent hydrophobicity.The affinity column without space arm was used to determine the human immunoglobulin G (HIgG) in human serum.The correlative coefficient of the calibration curve was 0.999.The effect of the flow rate on the pressure of the continuous rods was also investigated.The subclasses of HIgG were separated by the use of a pH gradient.
Keywords：Affinity chromatography,polymer,monolithic,protein A,human immunoglobulin G

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收稿日期：2000年6月20日

修稿日期：2000年12月28日

出版日期：2001年5月20日