Intact Proteoform Analysis by Capillary Electrophoresis–Mass Spectrometry. Are We There Yet?
Year: 2025
Type: article
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.
Authors: Noah Gould, Qianjie Wang, Jeffrey N. Agar, Jennifer S. Brodbelt, Daoyang Chen, Kellye A. Cupp‐Sutton, Elena Domínguez‐Vega, Fei Fang, Marianne Fillet, Matthew S Fischer, Attila Gaspar, Ying Ge, Marie‐Jia Gou, Michal Greguš, Christoph Gstöttner, Narmin Hamidli, Amanda Helms, Md Amin Hossain, Kyle J. Juetten, Neil L. Kelleher, Tobias Kraus, Eli J. Larson, J. Scott Mellors, Cynthia Nagy, Christian Neusüß, Erin A. Redman, Jasmin Schairer, Si Wu, Tian Xu, Zhitao Zhao, Guijie Zhu, Alexander R. Ivanov, Kevin Jooß, Liangliang Sun
Institutions: Northeastern University, Michigan State University, The University of Texas at Austin, Merck & Co., Inc., Rahway, NJ, USA (United States), University of Alabama, Leiden University Medical Center, University of Liège, Centre Hospitalier Universitaire de Liège, University of Wisconsin–Madison, University of Debrecen, Brigham and Women's Hospital, Harvard University, Northwestern University, Chemistry of Life Processes Institute, Hochschule Aalen, University of Oklahoma, Vrije Universiteit Amsterdam
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: hybrid
Estimated APC paid: $5,510
Funders: Oklahoma Center for the Advancement of Science and Technology, Universiteit Leiden, Leids Universitair Medisch Centrum, Nemzeti Kutatási Fejlesztési és Innovációs Hivatal, National Institutes of Health, National Research, Development and Innovation Office, National Heart, Lung, and Blood Institute, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases
Awards: G1061158614, G1451896423, G192239701, G2074959885, G3037769621, G3107205511, G3158393935, G3206484946, G3475589980, G3644809225, G4916275747, G5515374730, G582323035, G6838740781, G8525819481, G8701409497, G9000025781, G9000179385, G9000191144, G9000207411, G9000230575, G9000264098, G9000355098
OpenAlex ID: W4417456195