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J Proteome Res. 2012 Oct 5;11(10):5059-64. doi: 10.1021/pr300638n. Epub 2012 Sep 14.

Online nanoscale ERLIC-MS outperforms RPLC-MS for shotgun proteomics in complex mixtures.

Author information

1
Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, United States.

Abstract

We have explored the use of electrostatic repulsion hydrophilic interaction chromatography (ERLIC) as an alternative to the gold-standard in shotgun proteomics: reversed-phase (RP) LC for online ESI-MS/MS. Conditions for sample solubilization and initial gradient conditions were optimized to strike a balance between peptide solubility and maximum peptide retention when using mobile phase with high organic solvent concentration. Online ERLIC-MS demonstrated a 57% increase in total peptide identifications compared to RP-MS. We examined the mechanism of this improved performance and found that it stems from ERLIC's propensity to retain longer peptides, which can be identified with greater confidence. Online nanoscale ERLIC-MS provides a powerful new tool for enhancing MS-based shotgun proteomic in a broad range of applications.

PMID:
22950739
PMCID:
PMC3487405
DOI:
10.1021/pr300638n
[Indexed for MEDLINE]
Free PMC Article

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