Fractionation of proteins by immobilized metal affinity chromatography.

Abstract

It is widely known that the progress of proteomics mostly depends on the development of more advanced and sensitive protein separation technologies. Immobilized metal affinity chromatography (IMAC) is a useful protein fractionation method used to enrich metal associated proteins. IMAC represents an affinity separation approach based on the interaction between proteins and metal ions immobilized on a solid support. By changing various metal ions and other experimental conditions such as pH and elution composition, IMAC can selectively isolate metal-binding protein fractions for further specific proteomic analysis. The combination of IMAC with other protein analytical technologies has been successfully applied in characterizing metalloproteome and post-translational modifications. In the future, newly developed IMAC techniques integrated with other proteomic methods will significantly contribute to the further development of proteomics.