Immobilized Iron(III) Metal Affinity Chromatography for the Separation of Phosphorylated Macromolecules: Ligands and Applications

Abstract
This review highlights the advances in Fe(III)-Immobilized Metal Affinity Chromatography [Fe(III)-IMAC] for the separation and characterization of phosphorylated biomolecules. Fe(III)-IMAC has demonstrated a selective interaction with phosphate groups and other hard Lewis bases such as carboxylate and phenol functional groups. These properties make this technique useful for separating and studying macromolecules. Since phosphorylation is a central means of regulation in eukaryotes and prokaryotes, the development of Fe(III)-IMAC is important. Various iron chelating ligands and their properties are discussed. General features of Fe(III)-IMAC are examined and examples of useful separations are considered.