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化学学报
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化学学报 ›› 2010, Vol. 68 ›› Issue (23): 2421-2428. 上一篇    下一篇

研究论文

以原子转移自由基聚合法制备强阴离子交换色谱填料及其在蛋白质分离中的应用

王婀娜1，段淑静1，何锡文*,1，陈朗星1，李文友1，张玉奎*,1,2   

  1. (1南开大学化学学院 天津 300071)
    (2中国科学院大连化学物理研究所 大连 116011)
  • 投稿日期:2010-03-29 修回日期:2010-06-25 发布日期:2010-08-16
  • 通讯作者: 何锡文 E-mail:xiwenhe@nankai.edu.cn
  • 基金资助:

    国家重点基础研究发展规划(973计划);国家自然科学基金;国家自然科学基金

Preparation of anStrong Anion-Exchange Ion-Exchange Chromatographic Support by Atom Transfer Radical Polymerization and Their Application in pProtein s Separation

Wang Enauo1 Duan Shujing1 He Xiwen*,1 Chen Langxing1, Li Wenyou1 Zhang Yukui*,1,2   

  1. (1 Department of Chemistry, Nankai University, Tianjin 300071)
    (2 Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116011)
  • Received:2010-03-29 Revised:2010-06-25 Published:2010-08-16

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在大孔硅胶表面引入原子转移自由基聚合的引发基团, 通过该技术将甲基丙烯酰氧乙基三甲基氯化铵聚合于硅胶表面, 制得了强阴离子交换色谱填料. 详细考察了该填料对标准蛋白质的分离性能及流动相pH、流速、有机溶剂对蛋白质保留的影响. 实验结果表明, 在流速为0.75 mL/min时, 采用线性梯度洗脱, 20 分钟min内可快速分离不同体系的三种标准蛋白|并将该填料用于鸡蛋中卵清蛋白的快速分离纯化, 取得了很好的效果.

关键词: 硅胶, 强阴离子交换色谱, 原子转移自由基聚合, 蛋白质分离

A new strong anion-exchange stationary phase for HPLC has been synthesized by surface-initiated atom transfer radical polymerization (ATRP). Firstly, 2-bromoisobutyryl bromide as the ATRP initiator was attached onto the macroporous silica via chemical reaction. In the next stage, methacrylatoethyl trimethyl ammonium chloride (MATA) carrying strong anion exchanger groups as the monomer was polymerized on the initiator-immobilized particles via surface-initiated ATRP. After studying the adsorption properties, the suitable support was packed into stainless steel column to separate acid proteins by the linear gradient elution in 20 min at flow rate of 0.75 mL/min. The effects of flow rate, pH and organic solvents on the separation and retention ability of proteins were studied in detail. This support was also used for the separation and purification of ovalbumin from egg in only one step.

Key words: silica, strong anion-exchange chromatography, atom transfer radical polymerization, protein s separation