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Over 10 000 Peptide Identifications from the HeLa Proteome by Using Single‐Shot Capillary Zone Electrophoresis Combined with Tandem Mass Spectrometry
Author(s) -
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.
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201409075
Subject(s) - chemistry , chromatography , mass spectrometry , orbitrap , tandem mass spectrometry , proteome , capillary electrophoresis , tandem mass tag , shotgun proteomics , analytical chemistry (journal) , quantitative proteomics , isobaric labeling , protein mass spectrometry , proteomics , biochemistry , gene
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.