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.

metal chelate affinity chromatography; carboxymethylated aspartate ligand; histidine-tagged protein;