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
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  • 北京理工大学生命学院 北京 100081

收稿日期: 2011-08-11

  修回日期: 2011-11-08

  网络出版日期: 2012-03-17

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
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  • School of Life Science, Beijing Institute of Technology, Beijing 100081

Received date: 2011-08-11

  Revised date: 2011-11-08

  Online 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).

摘要

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.

本文引用格式

梁笑 , 刘宗建 , 戴荣继 , 王维 , 耿芳芳 , 梁艳丽 , 孟薇薇 , 邓玉林 . 肝素温度敏感色谱填料的制备与性能研究[J]. 化学学报, 2012 , 70(05) : 561 -566 . DOI: 10.6023/A1108112

Abstract

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.

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