Over 10000 Peptide Identifications from the HeLa Proteome by Using Single-Shot Capillary Zone Electrophoresis Combined with Tandem Mass Spectrometry

• Published in: Angewandte Chemie International Edition Vol. 53; no. 50; pp. 13931 - 13933
• Main Authors: Sun, Liangliang, Hebert, Alexander S, Yan, Xiaojing, Zhao, Yimeng, Westphall, Michael S, Rush, Matthew J P, Zhu, Guijie, Champion, Matthew M, Coon, Joshua J, Dovichi, Norman J
• Format: Journal Article
• Language: English
• Published: Weinheim Wiley Subscription Services, Inc 08.12.2014
• Edition: International ed. in English
• Subjects: Capillarity; Chromatography; Electrokinetics; Electrophoresis; Mass spectrometers; Mass spectrometry; Peptides; Proteins; Scientific imaging; State of the art
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/anie.201409075

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 90min) 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 10000 peptides and 2100 proteins from a HeLa cell proteome digest in approximately 100min. 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.