Mixed-mode hydrophilic interaction/cation-exchange chromatography (HILIC/CEX) of peptides and proteins

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 widel...

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Bibliographic Details
Published in:Journal of separation science 2008-08, Vol.31 (15), p.2754-2773
Main Authors: Mant, Colin T, Hodges, Robert S
Format: Article
Language:English
Subjects:
Analytical chemistry
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Cation Exchange Resins - chemistry
Chemistry
Chromatographic methods and physical methods associated with chromatography
Chromatography, Ion Exchange - methods
Exact sciences and technology
Fundamental and applied biological sciences. Psychology
General aspects, investigation methods
HILIC/CEX
Histones - analysis
Hydrophobic and Hydrophilic Interactions
Mixed-mode
Other chromatographic methods
Peptides
Peptides - analysis
Peptides - chemical synthesis
Proteins
Proteins - analysis
RPC
Sensitivity and Specificity
Time Factors
Water - chemistry
Wettability
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Summary: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, α-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.
ISSN:1615-9306
1615-9314
DOI:10.1002/jssc.200800243