4种金属离子亲和层析纯化重组类人胶原蛋白的效果比较  

Comparation of purified recombinant human-like collagen by affinity chromatography with four metal ions

作　　者：王晓军[1] 范代娣[2] 

机构地区：[1]西安工程大学环化学院,陕西西安710048 [2]西北大学化工学院,陕西西安710069

出　　处：《应用化工》2009年第6期857-859,共3页Applied Chemical Industry

摘　　要：对利用金属离子亲和层析纯化重组类人胶原蛋白过程中使用的金属离子进行了比较,从而对分离纯化的条件进行优化。在相同实验条件下,用4种金属离子柱分离纯化目的蛋白。结果显示,经4种金属离子柱纯化后镍柱的总蛋白收获率最高,铜柱与锌柱居中,钙柱最低;而柱保留时间则为锌柱最高(12.38 m in),钙柱最低(8.25 m in);钙柱洗脱时所需咪唑解离初始浓度最低(100 mmol/L),锌柱最高(200 mmol/L);对SDS-PAGE电泳图进行分析得纯化后类人胶原蛋白的纯度分别为:镍柱82.8%,铜柱83.4%,锌柱96.2%,钙柱94.3%。由此可见锌柱对目标蛋白的亲和力最高,且纯化后类人胶原蛋白的纯度也最高。因此,确定锌离子作为亲和层析纯化重组类人胶原蛋白的金属离子。Comparation of purified recominat human-like collagen by affinity chromatography. Then isddlion and puritication was optimizated. The target protein was purified by four metal columns under the same chromatographic conditions. The results showed that the protein harvest rate was the highest in the Ni column and the lowest in the Ca column. The retention time was the longest in Zn column （12.38 min）and the shortest in Ca column （8.25 min）. The initial concentration of imidazole was the highest in Zn column （200 mmol/L） and the lowest in Ca column （ 100 mmol/L）. By analysis of SDSPAGE electrophoresis gel it could be demonstrated that the collagen purity was 82.8% in Ni column, 83.4%in Cu column, 96.2% in Zn column and 94.3% in Ca column. It could be concluded that the affinity ability was the best in Zn column and purity of recombinant human-like collagen were the highest too. So the Zn ion was chosen to be the metal ion of IMAC for separation the recombinant human-like collagen.

关 键 词：重组类人胶原蛋白 金属螯合亲和层析 金属离子 纯化 

分 类 号：O636[理学—高分子化学]