Intact Proteoform Analysis by Capillary Electrophoresis–Mass Spectrometry. Are We There Yet?

Mass spectrometry (MS)-based top-down proteomics (TDP) has emerged as a powerful tool for characterizing proteoforms to advance both fundamental and translational research. TDP requires high-efficiency liquid-phase separation, high-resolution MS, and tandem MS. Capillary zone electrophoresis (CZE)-MS has been proposed as a promising analytical technique for protein analysis decades ago because of its unique and valuable features, including high separation efficiency and high detection sensitivity. However, CZE-MS has not been widely adopted by the proteomics community, mainly due to concerns with its robustness and reproducibility. Here, we hypothesized that CZE-MS is sufficiently robust and reproducible for broad adoption due to the continued efforts of the community over the last three decades. In this work, for the first time, research teams from around the world validated the robustness, repeatability, and reproducibility of CZE-MS for TDP in both simple and complex model proteoform mixtures employing a full spectrum of commercially available capillary electrophoresis (CE)-MS interfaces, instrumentation, and compared CZE-MS performance with state-of-the-art liquid chromatography (LC)-MS methods. This study offers the research community an informative resource of ready-to-use experimental CE-MS techniques and a better understanding of the CZE-MS approach and its potential in TDP, accelerating the broad adoption of CZE-MS in proteoform research.

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Publication Details

Journal
Angewandte Chemie International Edition
Published
2025-12-18
DOI
https://doi.org/10.1002/anie.202518366
Citations
2
Primary Topic
Advanced Proteomics Techniques and Applications
Type
article
Field-Weighted Citation Impact
1.51
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article

Intact Proteoform Analysis by Capillary Electrophoresis–Mass Spectrometry. Are We There Yet?

Jasmin Schairer, J. Scott Mellors, Eli J. Larson, Marianne Fillet et al.
2 citations
Angewandte Chemie International Edition
Advanced Proteomics Techniques and Applications
1.51
article

Intact Proteoform Analysis by Capillary Electrophoresis–Mass Spectrometry. Are We There Yet?

Jasmin Schairer, J. Scott Mellors, Eli J. Larson, Marianne Fillet, Attila Gáspár, Tobias Kraus, Matthew S. Fischer, Michal Greguš, Marie‐Jia Gou, Tian Xu, Cynthia Nagy, Si Wu, Qianjie Wang, Elena Domínguez‐Vega, Jeffrey N. Agar, Kevin Jooß, Amanda Helms, Christoph Gstöttner, Alexander R. Ivanov, Narmin Hamidli, Ying Ge, Kellye A. Cupp‐Sutton, Neil L. Kelleher, Fei Fang, Daoyang Chen, Kyle J. Juetten, Christian Neusüß, Noah Gould, Guijie Zhu, Zhitao Zhao, Md Amin Hossain, Jennifer S. Brodbelt, Liangliang Sun, Erin A. Redman
article en
2 citations

Abstract

Mass spectrometry (MS)-based top-down proteomics (TDP) has emerged as a powerful tool for characterizing proteoforms to advance both fundamental and translational research. TDP requires high-efficiency liquid-phase separation, high-resolution MS, and tandem MS. Capillary zone electrophoresis (CZE)-MS has been proposed as a promising analytical technique for protein analysis decades ago because of its unique and valuable features, including high separation efficiency and high detection sensitivity. However, CZE-MS has not been widely adopted by the proteomics community, mainly due to concerns with its robustness and reproducibility. Here, we hypothesized that CZE-MS is sufficiently robust and reproducible for broad adoption due to the continued efforts of the community over the last three decades. In this work, for the first time, research teams from around the world validated the robustness, repeatability, and reproducibility of CZE-MS for TDP in both simple and complex model proteoform mixtures employing a full spectrum of commercially available capillary electrophoresis (CE)-MS interfaces, instrumentation, and compared CZE-MS performance with state-of-the-art liquid chromatography (LC)-MS methods. This study offers the research community an informative resource of ready-to-use experimental CE-MS techniques and a better understanding of the CZE-MS approach and its potential in TDP, accelerating the broad adoption of CZE-MS in proteoform research.

Angewandte Chemie International EditionVol. 65(5)
Northwestern University (US), Analog Devices (United States) (US), Brigham and Women's Hospital (US), Merck & Co., Inc., Rahway, NJ, USA (United States) (US), Northeastern University (US), University of Debrecen (HU), University of Wisconsin–Madison (US), Harvard University (US), University of Liège (BE), University of Alabama (US), Leiden University Medical Center (NL), Academic Center for Dentistry Amsterdam (NL), Netherlands Metabolomics Centre (NL), Physical Devices (United States) (US), Merck Serono S.A. (Switzerland) (CH), Zero to Three (US), Hochschule Aalen (DE), Eastern University (BD), The University of Texas at Austin (US), Vrije Universiteit Amsterdam (NL), University of Oklahoma (US), Michigan State University (US)
Openalex Percentile: Top 15%
Advanced Proteomics Techniques and Applications
1.51
DOIOpen Access PDFOpenAlex
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