肝素温度敏感色谱填料的制备与性能研究  

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

作　　者：梁笑[1] 刘宗建[1] 戴荣继[1] 王维[1] 耿芳芳[1] 梁艳丽[1] 孟薇薇[1] 邓玉林[1] 

机构地区：[1]北京理工大学生命学院,北京100081

出　　处：《化学学报》2012年第5期561-566,共6页Acta Chimica Sinica

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

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.The silica beads modified by poly（N-isopropylacrylamide-co-N-acryloxysuccinimide） and hepa- rin, 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 tempera- ture-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 pro- vided a fast and efficient method for protein separation.

关 键 词：高效液相色谱 聚(N-异丙基丙烯酰胺) 肝素 凝血酶 

分 类 号：O657.7[理学—分析化学]