离子交换层析纯化重组福安泰-03功能域的工艺条件研究

Purification of Recombinant Functional Domain of Fuantai-03 by Ion Exchange Chromatography

对阴离子交换树脂HiTrap DEAE Sepharose FF纯化重组福安泰-03功能域(rFAT-03)的合适条件进行了研究。通过试验,确定了柱层析的最佳操作条件,即在上样缓冲液为0.1mol/L Tris-HCl、洗脱缓冲液为1mol/L NaCl+0.1mol/L Tris-HCl、pH为8.8的条件下,利用HiTrap DEAE SepharoseFF琼脂糖凝胶吸附柱,先用洗脱缓冲液从30%到60%洗脱3个柱床体积,再用洗脱缓冲液从60%到100%洗脱2个柱床体积。在此试验条件下,rFAT-03得率为35.97%,最终纯化的rFAT-03达电泳纯,纯度为99.5%。

The research the optimal conditions of purification of recombinant functional domain of Fuantai-03 was conducted by anion exchange resin HiTrap DEAE Sepharose FF. pH 8.8,0.1mol/L Tris-HCl was adsorption buffer system,and pH 8.8,0.1mol/L Tris-HCl,1mol/LNaCl was elution buffer system,elution buffer of eluent concentration is from 30% to 60% (3 column bed volume),and then elution buffer of eluent concentration was from 60% to 100% (2 column bed volume). In this experimental condition,recombinant functional domain of Fuantai-03 yield is 35.97%,and its purity is 99.5%.