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.%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.
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