title: Online nanoscale ERLIC-MS outperforms RPLC-MS for shotgun proteomics in complex mixtures.
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.
authors.affiliation: Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, United States.
authors.forename: Ebbing P
authors.initials: EP
authors.lastname: de Jong
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Griffin
mesh_terms: D000097:Acetonitriles / Q000737:chemistry; D002021:Buffers; D056148:Chromatography, Reverse-Phase; D006367:HeLa Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D036103:Nanotechnology; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012995:Solubility; D012997:Solvents / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D000097:Acetonitriles; D002021:Buffers; D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins; D012997:Solvents; C032159:acetonitrile