肝素温度敏感色谱填料的制备与性能研究

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.

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

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.