Analytical methods; Electrophoresis; Mass spectrometry; Proteomics; Proteins; Electrophoresis, Capillary/methods; Mass Spectrometry/methods; Proteomics/methods; Proteins/analysis; Analytical method; Capillary electrophoresis-mass spectrometries; Capillary-zone electrophoresis; Fundamental research; Higher efficiency; Liquid-phase separations; Reproducibilities; Top down proteomics; Translational Research; Electrophoresis, Capillary; Catalysis; Chemistry (all)
Abstract :
[en] 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.
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Gould, Noah ; Barnett Institute of Chemical and Biological Analysis and Department of Chemistry & Chemical Biology, Northeastern University, Boston, MA, USA
Wang, Qianjie; Department of Chemistry, Michigan State University, East Lansing, MI, USA
Agar, Jeffrey N; Barnett Institute of Chemical and Biological Analysis and Department of Chemistry & Chemical Biology, Northeastern University, Boston, MA, USA
Brodbelt, Jennifer S; Department of Chemistry, University of Texas, Austin, TX, USA
Chen, Daoyang; Discovery Biologics, Merck Research Laboratory, USA
Cupp-Sutton, Kellye A; Department of Chemistry and Biochemistry, University of Alabama, Tuscaloosa, AL, USA
Domínguez-Vega, Elena; Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands
Fang, Fei; Department of Chemistry, Michigan State University, East Lansing, MI, USA
Fillet, Marianne ; Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
Fischer, Matthew S; Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, 53706, USA
Gaspar, Attila; Department of Inorganic and Analytical Chemistry, University of Debrecen, Debrecen, Hungary
Ge, Ying; Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, 53706, USA ; Department of Cell and Regenerative Biology and Human Proteomics Program, University of Wisconsin-Madison, Madison, Wisconsin, 53705, USA
Gou, Marie-Jia; Laboratory for the Analysis of Medicines, Center for Interdisciplinary Research on Medicines (CIRM), University of Liege, Quartier Hospital, Liege, Belgium
Greguš, Michal; Barnett Institute of Chemical and Biological Analysis and Department of Chemistry & Chemical Biology, Northeastern University, Boston, MA, USA
Gstöttner, Christoph; Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands
Hamidli, Narmin; Department of Inorganic and Analytical Chemistry, University of Debrecen, Debrecen, Hungary
Helms, Amanda; Department of Chemistry, University of Texas, Austin, TX, USA
Hossain, Md Amin; Barnett Institute of Chemical and Biological Analysis and Department of Chemistry & Chemical Biology, Northeastern University, Boston, MA, USA ; Department of Neurosurgery, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA
Juetten, Kyle J; Department of Chemistry, University of Texas, Austin, TX, USA
Kelleher, Neil L; Departments of Chemistry and Molecular Biosciences, the Chemistry of Life Processes Institute, and the Proteomics Center of Excellence, Northwestern University, Evanston, IL, USA
Kraus, Tobias; Department of Chemistry, Aalen University, Aalen, Germany
Larson, Eli J; Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, 53706, USA
Mellors, J Scott; 908 Devices Inc., Boston, MA, USA
Nagy, Cynthia; Department of Inorganic and Analytical Chemistry, University of Debrecen, Debrecen, Hungary
Neusüß, Christian; Department of Chemistry, Aalen University, Aalen, Germany
Redman, Erin A; 908 Devices Inc., Boston, MA, USA
Schairer, Jasmin; Department of Chemistry, Aalen University, Aalen, Germany
Wu, Si; Department of Chemistry and Biochemistry, University of Alabama, Tuscaloosa, AL, USA ; Department of Chemistry and Biochemistry, University of Oklahoma, Norman, OK, USA
Xu, Tian; Departments of Chemistry and Molecular Biosciences, the Chemistry of Life Processes Institute, and the Proteomics Center of Excellence, Northwestern University, Evanston, IL, USA
Zhao, Zhitao; Department of Chemistry and Biochemistry, University of Alabama, Tuscaloosa, AL, USA ; Department of Chemistry and Biochemistry, University of Oklahoma, Norman, OK, USA
Zhu, Guijie; Department of Chemistry, Michigan State University, East Lansing, MI, USA
Ivanov, Alexander R ; Barnett Institute of Chemical and Biological Analysis and Department of Chemistry & Chemical Biology, Northeastern University, Boston, MA, USA
Jooß, Kevin; 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
Sun, Liangliang; Department of Chemistry, Michigan State University, East Lansing, MI, USA
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