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化学学报
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化学学报 ›› 2012, Vol. 70 ›› Issue (05): 561-566 .DOI: 10.6023/A1108112 上一篇    下一篇

研究论文

肝素温度敏感色谱填料的制备与性能研究

梁笑, 刘宗建, 戴荣继, 王维, 耿芳芳, 梁艳丽, 孟薇薇, 邓玉林   

  1. 北京理工大学生命学院 北京 100081
  • 投稿日期:2011-08-11 修回日期:2011-11-08 发布日期:2012-03-17
  • 通讯作者: 戴荣继, 邓玉林 E-mail:dairongji@bit.edu.cn; Deng@bit.edu.cn
  • 基金资助:

    中国国家自然科学基金(No. 20675008)和中国自然基础研究发展计划(No. 2007CB914103)资助项目.

Preparation and Characterization of Temperature-responsive Affinity Chromatographic Stationary Phase Modified with Heparin

Liang Xiao, Liu Zongjian, Dai Rongji, Wang Wei, Geng Fangfang, Liang Yanli, Meng Weiwei, Deng Yulin   

  1. School of Life Science, Beijing Institute of Technology, Beijing 100081
  • Received:2011-08-11 Revised:2011-11-08 Published:2012-03-17
  • Supported by:

    Project supported by the National Natural Science Foundation of China (No. 20675008) and the National Basic Research Program of China (973 Program, No. 2007CB914103).

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利用自由基聚合法制备了含有聚(N-异丙基丙烯酰胺/N-丙烯酰氧琥珀酰亚胺)和肝素共聚物的温度敏感的亲和色谱填料; 通过热失重分析得到该填料表面聚合物的接枝率为9.45%; Elison-Morgan 法测定该填料表面的肝素含量为3.6mg/g. 将其用作HPLC 固定相, 分离苯和氢化可的松, 表明该色谱固定相具有温度敏感特性; 该色谱柱对凝血酶具有较强的亲和作用, 通过控制温度缩短了凝血酶的释放时间, 回收率为81.4%, 这为蛋白质的分离提供了一个快速有效的方法.

关键词: 高效液相色谱, 聚(N-异丙基丙烯酰胺), 肝素, 凝血酶

The silica beads modified by poly(N-isopropylacrylamide-co-N-acryloxysuccinimide) and heparin, which was then used as temperature-responsive affinity chromatographic stationary phase were prepared via radical polymerization. Grafting ratio of polymers grafted on surfaces was 9.45% calculated by thermogravimetric analysis (TGA) and the heparin content on surfaces was 3.6 mg/g determined by Elson-Morgan method. The packing column with the modified silica beads as the HPLC stationary phase had temperature- responsive features, which was proved by separating benzene and hydrocortisone. The results also indicated that the strong affinity interaction occurs between thrombin and the modified column, with recovery of 81.4%. The releasing time of thrombin was shortened by controlling the column temperature, which provided a fast and efficient method for protein separation.

Key words: high-performance liquid chromatography, poly(N-isopropylacrylamide), heparin, thrombin