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将KGM凝胶和Sepharose 4B在同样条件下活化偶联,制成Cu~(2+)金属螫合亲和胶,亲和纯化猪血SOD,并对这两种亲和胶的层析效果和性能进行了比较。KGM金属螫合胶对猪血SOD吸附量、纯化倍数、纯化SOD的比活力和回收率分别为53000U/ml胶、19倍、12000U/mg蛋白和94.6％,而Sepharose 4B亲和胶对SOD 的吸附量、纯化倍数、纯化SOD的比活力和回收率分别为79920U/ml胶、11倍、10125U/mg蛋白和95.4％。两种亲和胶所纯化的SOD经聚丙烯酰胺凝胶电泳(PAGE)、活性染色及SDS聚丙烯酰胺凝胶电泳(SDS-PAGE)证明其均为电泳纯。KGM金属螯合胶使用六次后,其对SOD吸附量、去Cu量及SOD的回收率均无明显影响。
KGM gel and Sepharose 4B were activated and coupled under the same conditions to make Cu 2+ metal affinity gel and affinity purification of porcine blood SOD. The chromatographic effect of these two kinds of affinity gel and Performance is compared. The specific activity and the recoveries of KGM metal chelated gum on pig blood SOD adsorption amount, purification fold, purified SOD were 53000U / ml, 19 times, 12000U / mg protein and 94.6%, respectively, while that of Sepharose 4B affinity resin on SOD The specific activity and recovery of purified SOD were 79920U / ml, 11X, 10125U / mg and 95.4% respectively. SOD purified by two kinds of affinity gums was proved to be electrophoretic pure by polyacrylamide gel electrophoresis (PAGE), active dyeing and SDS polyacrylamide gel electrophoresis (SDS-PAGE). After using KGM metal chelate gel six times, the adsorption capacity of SOD, the amount of Cu removal and the recovery rate of SOD had no significant effect.