A procedure for the preparation of continuous rod consisting of poly(glycidyl methacrylate\|co\|ethylene dimethacrylate) for immobilized metal affinity chromatography (IMAC) is presented. When chelating Cu(Ⅱ), Zn(Ⅱ)...A procedure for the preparation of continuous rod consisting of poly(glycidyl methacrylate\|co\|ethylene dimethacrylate) for immobilized metal affinity chromatography (IMAC) is presented. When chelating Cu(Ⅱ), Zn(Ⅱ) or Ni(Ⅱ) on it, the rod displayed a property of IMAC and the selectivity for protein separation was different from that obtained from the naked rod. An abnormal increase in retention times of proteins was found under the condition of either very high pH or very low pH on the metal chelated rods.展开更多
An immobilized metal ion affinity membrane was prepared by plasma-induced graft polymerization of glycidyl methacrylate (GMA) onto a porous polypropylene(PP), followed by chemical conversion of epoxide group of the GM...An immobilized metal ion affinity membrane was prepared by plasma-induced graft polymerization of glycidyl methacrylate (GMA) onto a porous polypropylene(PP), followed by chemical conversion of epoxide group of the GMA grafted membrane into an iminodiacetate(IDA) group, and chelation with copper ion. The effect of plasma discharge condition, GMA monomer concentration and grafting time on the degree of GMA grafting was studied and the optimum condition for plasma treatment of membrane was:10 W for 30 s, and that for grafting is 30 h at a concentration of 1 mol· L -1 GMA.Under this condition, a maximum grafting degree of 13.28% was obtained. The spectra of IR and XPS shows that GMA and IDA had been attached to the PP membrane after grafting and coupling. The nonselective adsorption of PP membrane had been significantly reduced after the H 2SO 4 treatment of the IDA coupled membrane. Finally, the isotherm adsorption of bovine serum albumin (BSA) on the immobilized metal ion affinity membrane was measured and the adsorption capacity of BSA increased with the degree of grafting. When BSA concentration was 1080 μg·ml -1 ,the adsorption ability of the affinity membrane with a grafting degree of 5.3% was nearly twice as much as that membrane with a grafting degree of 4.66%.展开更多
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...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。展开更多
文摘A procedure for the preparation of continuous rod consisting of poly(glycidyl methacrylate\|co\|ethylene dimethacrylate) for immobilized metal affinity chromatography (IMAC) is presented. When chelating Cu(Ⅱ), Zn(Ⅱ) or Ni(Ⅱ) on it, the rod displayed a property of IMAC and the selectivity for protein separation was different from that obtained from the naked rod. An abnormal increase in retention times of proteins was found under the condition of either very high pH or very low pH on the metal chelated rods.
文摘An immobilized metal ion affinity membrane was prepared by plasma-induced graft polymerization of glycidyl methacrylate (GMA) onto a porous polypropylene(PP), followed by chemical conversion of epoxide group of the GMA grafted membrane into an iminodiacetate(IDA) group, and chelation with copper ion. The effect of plasma discharge condition, GMA monomer concentration and grafting time on the degree of GMA grafting was studied and the optimum condition for plasma treatment of membrane was:10 W for 30 s, and that for grafting is 30 h at a concentration of 1 mol· L -1 GMA.Under this condition, a maximum grafting degree of 13.28% was obtained. The spectra of IR and XPS shows that GMA and IDA had been attached to the PP membrane after grafting and coupling. The nonselective adsorption of PP membrane had been significantly reduced after the H 2SO 4 treatment of the IDA coupled membrane. Finally, the isotherm adsorption of bovine serum albumin (BSA) on the immobilized metal ion affinity membrane was measured and the adsorption capacity of BSA increased with the degree of grafting. When BSA concentration was 1080 μg·ml -1 ,the adsorption ability of the affinity membrane with a grafting degree of 5.3% was nearly twice as much as that membrane with a grafting degree of 4.66%.
文摘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。
