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 language | English (US) |
---|---|
Pages (from-to) | 5059-5064 |
Number of pages | 6 |
Journal | Journal of Proteome Research |
Volume | 11 |
Issue number | 10 |
DOIs | |
State | Published - Oct 5 2012 |
Keywords
- mass spectrometry
- online ERLIC
- reversed phase liquid chromatography
- shotgun proteomics