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

Ebbing P. De Jong, Timothy J. Griffin

Research output: Contribution to journalArticle

18 Scopus citations

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 ERLICs 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.

Original languageEnglish (US)
Pages (from-to)5059-5064
Number of pages6
JournalJournal of Proteome Research
Volume11
Issue number10
DOIs
StatePublished - Oct 5 2012

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Keywords

  • mass spectrometry
  • online ERLIC
  • reversed phase liquid chromatography
  • shotgun proteomics

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