色谱

色谱

• 研究报告 •    下一篇

单分散树脂基质的弱阳离子交换色谱固定相的制备及其在生物大分子分离中的应用研究

李龙1,马桂娟1,龚波林1,2

  

  1. 1. Key Laboratory of Energy &Chemical Engineering, Ningxia University, Yinchuan 750021, China;
    2. Key Laboratory of Biotechnology, Ningxia University, Yinchuan 750021, China
  • 收稿日期:2005-07-01 修回日期:2005-08-31 出版日期:2005-11-30 发布日期:1987-12-25
  • 通讯作者: 龚波林

Preparation of Weak Cation-Exchange Packings Based on Monodisperse Porous Poly (Chloromethylstyrene-co-
Divinylbenzene) Beads and Its Applications in Separation of Biopolymers

LI Long1, MA Guijuan1, GONG Bolin1,2   

  1. 1. Key Laboratory of Energy &Chemical Engineering, Ningxia University, Yinchuan 750021, China;
    2. Key Laboratory of Biotechnology, Ningxia University, Yinchuan 750021, China
  • Received:2005-07-01 Revised:2005-08-31 Online:2005-11-30 Published:1987-12-25

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摘要： 采用分散聚合法制备种子和“一步种子溶胀聚合法”制备了粒径为6~15 μm的单分散多孔氯甲基苯乙烯-二乙烯苯微球。该微球经化学改性后得到一种亲水性良好的新型高效弱阳离子交换色谱固定相。详细考察了该固定相的表面亲水性、对标准蛋白的分离性能和盐的种类对蛋白质保留行为的影响。考察结果表明该固定相是一种性能优异的弱阳离子交换色谱固定相。将其应用于鸡蛋清中溶菌酶的快速分离纯化,纯化后的溶菌酶纯度高于96%,比活高达71184 U/mg。

关键词: 单分散多孔交联氯甲基苯乙烯-二乙烯苯微球, 蛋白质分离 , 弱阳离子交换色谱

Abstract:

Monodisperse polystyrene seed particles were prepared by dispersion polymerization of styrene in ethanol and water under a nitrogen atmosphere using benzoyl peroxide as an initiator. Then the monodisperse, porous poly (chloromethylstyrene-co-divinylbenzene) beads in the range of 6-15 μm were prepared by one-step swelling and polymerization method. Based on this media, a new hydrophilic weak cation exchange stationary phase (WCX) for high performance liquid chromatography was synthesized by a new chemical modification method. The stationary phase was evaluated on the behavior of ion exchange, separability, and hydrophilicity. Effects of the nature of salts and pH value on the separation and retention of proteins of the stationary phase were investigated in details. It was found that the WCX resin follows ion exchange chromatographic retention mechanism. It was also used for the rapid separation and purification of lysozyme from egg white in only one step. The purity was found more than 96% and specific bioactivity of the purified lysozyme was as high as 71184 U/mg.

Key words: protein separation , weak cation exchange chromatography, monodisperse porous poly (chloromethylstyrene-co-divinylbenzene) beads