Hydrophobic Interaction Chromatography

Abstract

Hydrophobic interaction chromatography (HIC) involves the separation of protein molecules in their native and biologically active state owing to a differential interaction of these molecules with hydrophobic sites on the surface of a solid support. In the separation process, hydrophobic patches on the protein surface interact with hydrophobic molecules (e.g. alkyl residues) immobilised on the hydrophilic solid phase surface (e.g. agarose). In the paper, a historical overview is followed by a consideration of the biochemical and biophysical mechanisms involved in HIC and a discussion of the critical hydrophobicity method and other approaches to hydrophobic interaction chromatographic methods. A detailed protocol for the individual synthesis of gel libraries for specific needs has been published. The treatise is concluded with a discussion on the predictability of chromatographic results and an outlook into the future.