Intact Proteoform Analysis by Capillary Electrophoresis–Mass Spectrometry. Are We There Yet?
Abstract
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.
Article Details
Authors (34)
Noah Gould
Barnett Institute of Chemical and Biological Analysis and Department of Chemistry & Chemical Biology Northeastern University Boston MA USA
Qianjie Wang
Department of Chemistry Michigan State University East Lansing MI USA
Jeffrey N. Agar
Jennifer S. Brodbelt
Daoyang Chen
Discovery Biologics Merck Research Laboratory USA
Kellye A. Cupp‐Sutton
Department of Chemistry and Biochemistry University of Alabama Tuscaloosa AL USA
Elena Domínguez‐Vega
Center for Proteomics and Metabolomics Leiden University Medical Center Leiden The Netherlands
Fei Fang
Marianne Fillet
Laboratory for the Analysis of Medicines Center for Interdisciplinary Research on Medicines (CIRM) University of Liege Quartier Hospital Liege Belgium
Matthew S Fischer
Department of Chemistry University of Wisconsin‐Madison Madison Wisconsin 53706 USA
Attila Gáspár
Ying Ge
Marie‐Jia Gou
Laboratory for the Analysis of Medicines Center for Interdisciplinary Research on Medicines (CIRM) University of Liege Quartier Hospital Liege Belgium
Michal Greguš
Christoph Gstöttner
Center for Proteomics and Metabolomics Leiden University Medical Center Leiden The Netherlands
Narmin Hamidli
Department of Inorganic and Analytical Chemistry University of Debrecen Debrecen Hungary
Amanda Helms
Department of Chemistry, The University of Texas at Austin
Md Amin Hossain
Kyle J. Juetten
Department of Chemistry University of Texas Austin TX USA
Neil L. Kelleher
Department of Chemistry
Tobias Kraus
Department of Chemistry Aalen University Aalen Germany
Eli J. Larson
Department of Chemistry University of Wisconsin‐Madison Madison Wisconsin 53706 USA
J. Scott Mellors
908 Devices Inc. Boston MA USA
Cynthia Nagy
Department of Inorganic and Analytical Chemistry University of Debrecen Debrecen Hungary
Christian Neusüß
Department of Chemistry Aalen University Aalen Germany
Erin A. Redman
908 Devices Inc. Boston MA USA
Jasmin Schairer
Department of Chemistry Aalen University Aalen Germany
Si Wu
Tian Xu
Department of Chemical and Biomolecular Engineering and Institute for NanoBioTechnology, The Johns Hopkins University, 3400 North Charles Street, Baltimore, Maryland 21218, United States
Zhitao Zhao
Guijie Zhu
Department of Chemistry Michigan State University East Lansing MI USA
Alexander R. Ivanov
Barnett Institute of Chemical and Biological Analysis and Department of Chemistry & Chemical Biology Northeastern University Boston MA USA
Kevin Jooß
Division of Bioanalytical Chemistry Department of Chemistry and Pharmaceutical Sciences Amsterdam Institute of Molecular and Life Sciences Vrije Universiteit Amsterdam Centre for Analytical Sciences Amsterdam (CASA) Amsterdam The Netherlands
Liangliang Sun
Department of Chemistry, Michigan State University