title: Mixed-mode hydrophilic interaction/cation-exchange chromatography (HILIC/CEX) of peptides and proteins.
abstract: This review represents a summary of the development and application of a novel mixed-mode HPLC approach to the separation and analysis of peptides and proteins termed hydrophilic interaction/cation-exchange chromatography (HILIC/CEX). This approach combines the most advantageous aspects of two widely different separation mechanisms, i.e. a separation based on hydrophilicity/hydrophobicity differences between polypeptides overlaid on a separation based on net charge. Applications described include HILIC/CEX separations of cyclic peptides, alpha-helical peptides, random coil peptides and modified or deletion products of synthetic peptides. In addition, the excellent resolving ability of HILIC/CEX for modified histone proteins is described. This approach is shown to represent an excellent complement to RP chromatography (RPC), as well as being a potent analytical tool in its own right.
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Colorado Denver, School of Medicine, Anschutz Medical Campus, Aurora, CO 80045, USA.
authors.forename: Colin T
authors.initials: CT
authors.lastname: Mant
authors.forename: Robert S
authors.initials: RS
authors.lastname: Hodges
mesh_terms: D002411:Cation Exchange Resins / Q000737:chemistry*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006657:Histones / Q000032:analysis; D057927:Hydrophobic and Hydrophilic Interactions; D010455:Peptides / Q000032:analysis* / Q000138:chemical synthesis; D011506:Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014867:Water / Q000737:chemistry; D017155:Wettability
chemical_list: D002411:Cation Exchange Resins; D006657:Histones; D010455:Peptides; D011506:Proteins; D014867:Water