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Mixed‐mode hydrophilic interaction/cation‐exchange chromatography (HILIC/CEX) of peptides and proteins
Author(s) -
Mant Colin T.,
Hodges Robert S.
Publication year - 2008
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200800243
Subject(s) - hydrophilic interaction chromatography , chemistry , chromatography , high performance liquid chromatography
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.