Acta Chimica Sinica ›› 2012, Vol. 70 ›› Issue (05): 561-566 .DOI: 10.6023/A1108112 Previous Articles     Next Articles

Full Papers


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;
  • 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).

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