壳聚糖微珠亲和层析介质的制备及其初步应用

Preparation and Preliminary Application of Chitosan Beads for Affinity Chromatography

目的制备壳聚糖(CTS)微珠亲和层析介质,并进行初步应用。方法采用多种化学方法对CTS的表面基团进行改造,使其活化。用此改性CTS进行羊抗人IgG及小牛血清纤维连接蛋白(FN)的纯化。结果此改性的CTS微珠粒径在40￣160μm之间,每克可溶胀至4ml。活化的CTS对人丙种球蛋白的吸附量为15￣25mg/gCTS,对明胶的吸附量为5.8￣6.5mg/gCTS。采用此方法纯化的羊抗人IgG及FN与Sepharose4B法纯化效果一致。结论已制备的壳聚糖微珠亲和层析介质,其性质稳定且价格较低,是一种可以广泛应用的亲和层析介质。

Objective To prepare and preliminarily apply the chitosan （CTS） beads as a medium for affinity chromatography. Methods Activate CTS by modifying its surface groups with various chemical methods. Purify goat anti-human IgG and calf serum fibronectin with the modified CTS. Results The size of modified CTS beads ranged from 40 to 160 μm. Each gram of CTS beads could be swollen to a volume of 4 ml. The adsorption capacities of the activated CTS to human gamma globulin and gelatin were 15 - 25 and 5. 8 - 6.5 mg/g CTS respectively. Both the purification effects of goat anti-human IgG and calf serum fibronectin with the modified CTS were identical to that with Sepharose 4B. Conclusion The prepared CTS beads were stable and low-cost, and might be used widely for affinity chromatography.