title: Over 10,000 peptide identifications from the HeLa proteome by using single-shot capillary zone electrophoresis combined with tandem mass spectrometry.
abstract: Capillary zone electrophoresis (CZE)-tandem mass spectrometry (MS/MS) has recently attracted attention as a tool for shotgun proteomics. However, its performance for this analysis has so far fallen far below that of reversed-phase liquid chromatography (RPLC)-MS/MS. The use of a CZE method with a wide separation window (up to 90 min) and high peak capacity (ca. 300) is reported. This method was coupled to an Orbitrap Fusion mass spectrometer through an electrokinetically pumped sheath-flow interface for the analysis of complex proteome digests. Single-shot CZE-MS/MS lead to the identification of over 10 000 peptides and 2100 proteins from a HeLa cell proteome digest in approximately 100 min. This performance is nearly an order of magnitude better than earlier CZE studies and is within a factor of two to four of the state-of-the-art nano ultrahigh-pressure LC system.
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556 (USA).
authors.forename: Liangliang
authors.initials: L
authors.lastname: Sun
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Hebert
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Yan
authors.forename: Yimeng
authors.initials: Y
authors.lastname: Zhao
authors.forename: Michael S
authors.initials: MS
authors.lastname: Westphall
authors.forename: Matthew J P
authors.initials: MJ
authors.lastname: Rush
authors.forename: Guijie
authors.initials: G
authors.lastname: Zhu
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Champion
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Coon
authors.forename: Norman J
authors.initials: NJ
authors.lastname: Dovichi
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D010455:Peptides / Q000737:chemistry*; D020543:Proteome; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D010455:Peptides; D020543:Proteome
keywords: CZE-MS/MS; analytical methods; electrophoresis; mass spectrometry; proteomics