金属螯合亲和介质吸附重组类人胶原蛋白条件研究

Study On Metal-Chelated Affinity Adsorption Conditions For Ecombinant Human-Like Collagen

本研究对利用金属螯合亲和方法纯化重组类人胶原蛋白的吸附条件进行了优化。采用分批式操作对金属离子、平衡缓冲液的类型、pH值、离子强度及上样量进行了选择,结果显示当采用Cu离子进行螯合,pH7.5、含0.5mol·L^(-1)NaCl的磷酸盐缓冲液作为平衡缓冲液,上样量为1.5ml/m1介质时,吸附效果最佳。

Recombinant human-like collagen with a hexahistidine tail attacked to the carboxyl end could be purified by Metal-chelated affinity adsorption. To choose the best adorption conditions, the metal ion, equilibrate buffer, pH, ionic strength and the volume of sample were optimized by using the batch chromatography. The results were that Cu^2＋ as the metal ion, 20mM Na-phosphate buffer pH 7.5 containing 0.75M NaCI as the equilibrate buffer and 1.6ml sample per millilitre marx.