Mixed-mode hydrophilic interaction/cation-exchange chromatography (HILIC/CEX) of peptides and proteins |
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Authors: | Mant Colin T Hodges Robert S |
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Institution: | Department of Biochemistry and Molecular Genetics, University of Colorado Denver, School of Medicine, Anschutz Medical Campus, Aurora, CO 80045, USA. |
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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. |
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Keywords: | HILIC/CEX Mixed‐mode Peptides Proteins RPC |
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