直接偶联法制备配基密度可控的丁基琼脂糖层析介质

Preparation of Butyl-agarose Chromatography Media with Controlled Ligand Density through Direct Coupling Reaction

以快流速琼脂糖凝胶为基质,采用直接偶联法制备丁基琼脂糖疏水层析介质.考察了影响反应的主要因素,通过四因素三水平正交实验和单因素实验,确定丁基琼脂糖疏水层析介质的优化反应条件为:反应温度45℃,反应时间45min,催化剂三氟化硼乙醚用量0.02mL/g.在此条件下严格控制琼脂糖凝胶的含水量,改变丁基缩水甘油醚用量,制备了不同丁基密度的丁基琼脂糖疏水层析介质.用不同密度的系列介质考察对牛血清白蛋白的吸附性能,初步研究了配基密度与吸附性能的关系;研究了疏水层析介质的机械稳定性和化学稳定性.结果表明,其各项性能良好,具有广阔的应用前景.

Butyl-agarose chromatography media were prepared by direct coupling of butyl glycidyl ether （BGE） on agarose 4FF beads. The effects of various reaction parameters on the density of butyl group were investigated. The preparative conditions were optimized by erthogonal design of the experiments. The optimum conditions were found to be 0.02 mL/g （BF3·Et2O/gel）, reaction time 45 min and reaction temperature 45℃. The ligand density could be controlled by the amount of BGE added to the reaction mixture. The mechanical stability, chemical stability and adsorption capability were also investigated. The results demonstrated that the butyl-agarose chromatography media with varied ligand density would be promising media for protein separation and purification.