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

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 wi...

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Bibliographic Details
Published in:Angewandte Chemie 2014-12, Vol.126 (50), p.14151-14153
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: Article
Language:eng ; ger
Subjects:
Capillarity
Chemistry
Chromatography
Electrokinetics
Electrophoresis
Mass spectrometers
Mass spectrometry
Nanostructure
Peptides
Scientific imaging
State of the art
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Summary: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.
ISSN:0044-8249
1521-3757
DOI:10.1002/ange.201409075