Deep Top-Down Proteomics Using Capillary Zone Electrophoresis-Tandem Mass Spectrometry: Identification of 5700 Proteoforms from the <i>Escherichia coli</i> Proteome

McCool, Elijah N.; Lubeckyj, Rachele A.; Shen, Xiaojing; Chen, Daoyang; Kou, Qiang; Liu, Xiaowen; Sun, Liangliang

doi:10.1021/acs.analchem.8b00693

Cited by 102 publications

(122 citation statements)

References 39 publications

(74 reference statements)

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“…The group used a similar platform, replacing SCX with SEC for top‐down proteomics of the E. coli proteome. They were able to identify 5700 proteoforms by SEC‐RPLC pre‐fractioning combined with CZE‐MS .…”

Section: Analytical Applicationsmentioning

confidence: 99%

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

et al. 2018

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Capillary electrophoresis (CE) offers fast and high‐resolution separation of charged analytes from small injection volumes. Coupled to mass spectrometry (MS), it represents a powerful analytical technique providing (exact) mass information and enables molecular characterization based on fragmentation. Although hyphenation of CE and MS is not straightforward, much emphasis has been placed on enabling efficient ionization and user‐friendly coupling. Though several interfaces are now commercially available, research on more efficient and robust interfacing with nano‐electrospray ionization (ESI), matrix‐assisted laser desorption/ionization (MALDI) and inductively coupled plasma mass spectrometry (ICP) continues with considerable results. At the same time, CE‐MS has been used in many fields, predominantly for the analysis of proteins, peptides and metabolites. This review belongs to a series of regularly published articles, summarizing 248 articles covering the time between June 2016 and May 2018. Latest developments on hyphenation of CE with MS as well as instrumental developments such as two‐dimensional separation systems with MS detection are mentioned. Furthermore, applications of various CE‐modes including capillary zone electrophoresis (CZE), nonaqueous capillary electrophoresis (NACE), capillary gel electrophoresis (CGE) and capillary isoelectric focusing (CIEF) coupled to MS in biological, pharmaceutical and environmental research are summarized.

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Section: Analytical Applicationsmentioning

confidence: 99%

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

et al. 2018

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“…To facilitate the wide separation window and increased sample volume, a separation capillary with a high‐quality neutral coating on its inner wall and a highly efficient protein stacking method based on a dynamic pH junction principle were utilized. They also coupled SEC‐RPLC to CZE‐MS/MS and identified nearly 6000 proteoforms and 850 proteoform families, which is one of the largest reported top‐down proteomic datasets thus far . A detailed protocol for CZE‐MS/MS‐based top‐down proteomics has been provided to facilitate adoption of this technique .…”

Section: Separation Of Proteoformsmentioning

confidence: 99%

Identification and Quantification of Proteoforms by Mass Spectrometry

et al. 2019

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A proteoform is a defined form of a protein derived from a given gene with a specific amino acid sequence and localized post‐translational modifications. In top‐down proteomic analyses, proteoforms are identified and quantified through mass spectrometric analysis of intact proteins. Recent technological developments have enabled comprehensive proteoform analyses in complex samples, and an increasing number of laboratories are adopting top‐down proteomic workflows. In this review, some recent advances are outlined and current challenges and future directions for the field are discussed.

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“…Alternatively, McCool et al. presented an orthogonal multidimensional separation platform that couples SEC and RPLC based protein prefractionation to CZE–MS/MS for deep top‐down proteomics of E . coli.…”

Section: Stackingmentioning

confidence: 99%

Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2016–2018)

et al. 2018

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One of the most cited limitations of capillary and microchip electrophoresis is the poor sensitivity. This review continues to update this series of biannual reviews, first published in Electrophoresis in 2007, on developments in the field of online/in‐line concentration methods in capillaries and microchips, covering the period July 2016–June 2018. It includes developments in the field of stacking, covering all methods from field‐amplified sample stacking and large‐volume sample stacking, through to isotachophoresis, dynamic pH junction, and sweeping. Attention is also given to online or in‐line extraction methods that have been used for electrophoresis.

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Deep Top-Down Proteomics Using Capillary Zone Electrophoresis-Tandem Mass Spectrometry: Identification of 5700 Proteoforms from the Escherichia coli Proteome

Cited by 102 publications

References 39 publications

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

Identification and Quantification of Proteoforms by Mass Spectrometry

Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2016–2018)

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