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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Published in | Angewandte Chemie International Edition Vol. 53; no. 50; pp. 13931 - 13933 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
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Weinheim
Wiley Subscription Services, Inc
08.12.2014
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Edition | International ed. in English |
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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 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. |
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AbstractList | 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. 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. Let's see some ID: 2100 protein and 10000 peptide identifications (IDs) from a HeLa cell proteome digest were obtained in a single 100min capillary zone electrophoresis (CZE)-tandem mass spectrometry (MS/MS) analysis. These results represent an almost tenfold improvement in peptide and protein IDs compared with previous single-shot analyses using this technology. |
Author | Zhao, Yimeng Sun, Liangliang Dovichi, Norman J Coon, Joshua J Hebert, Alexander S Champion, Matthew M Westphall, Michael S Yan, Xiaojing Rush, Matthew J P Zhu, Guijie |
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SubjectTerms | Capillarity Chromatography Electrokinetics Electrophoresis Mass spectrometers Mass spectrometry Peptides Proteins Scientific imaging State of the art |
Title | Over 10000 Peptide Identifications from the HeLa Proteome by Using Single-Shot Capillary Zone Electrophoresis Combined with Tandem Mass Spectrometry |
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