【摘要】 以琼脂糖(agarose)凝胶的珠状产品SepharoseCl-6B为基质,以环氧氯丙烷为活化剂,将天冬氨酸固定在琼脂糖凝胶微球上,在经与溴乙酸修饰后,最终得到含有羧甲基天冬氨酸结构的琼脂糖凝胶微球.以凝胶微球负载的多羧基为配体分别与Co2+及Ni2+配位分别得到两种金属螯合亲和层析介质.依照上述方法制备了具有不同负载羧基含量的色谱介质.以六聚组氨酸融合蛋白为分离样品,研究了所制备的色谱分离介质的分离纯化性能,并与商品化色谱分离介质Agarose-NTA-Ni进行了比较.结果表明,目标介质蛋白结合容量大,与蛋白结合快、易洗脱、选择性高,金属离子不易脱落.更多还原

【Abstract】 The metal chelate affinity chromatographic materials were successfully prepared with Sepharose CL-6B as support,3-Chloro-1,2-epoxypropane as activated agent,carboxymethylated aspartate(CM-Asp) as chelating ligand and Co2+ and Ni2+ as center ions.The metal chelate affinity chromatographic materials were characterized using acid-base titration,IR,AAS,SEM and EDX,respectirely.The results indicate that metal ions are chelated to the carboxyl groups immobilized onto agarose to form multidentate complexes.The results from purification of 6×His recombination protein release that the efficiency in purification of tagged protein derived from the affinity chromatographic material containing Co2+ is higher than that containing Ni2+.Additionally,the prepared affinity chromatographic material containing Co2+ is more efficient in purification of 6×His recombination protein than a commercially available affinity chromatographic material,Agarose-NTA-Ni.更多还原