Preparation of metal chelate affinity chromatographic material and its application in purification of histidine-tagged protein

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.