Fast separation and analysis of reduced monoclonal antibodies with capillary zone electrophoresis coupled to mass spectrometry
Abstract
Capillary zone electrophoresis-electrospray ionization-mass spectrometry (CZE-ESI-MS) was used for analysis of reduced antibodies. We first developed a simple protocol to condition commercial linear-polyacrylamide coated capillaries for use in top-down proteomics. We then suspended reduced antibodies in a solution of 35% acetic acid, 50% acetonitrile in water. Heavy and light chains were baseline resolved within 10 min and with 3-30 µg/mL detection limits using a 0.1% aqueous formic acid background electrolyte. Quintuplicate runs of a two-antibody mixture produced relative standard deviations of ∼1% in migration time and 10% in peak amplitudes. Resolution was further improved for the two-antibody mixture by using 5% acetic acid as the background electrolyte, highlighting the potential of capillary electrophoresis-mass spectrometry for analysis of antibody mixtures.
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Notable CZE-MS examples include intact mass analysis of biopharmaceutical proteins and monoclonal antibodies, where complete baseline separation is often achieved.
“Notable examples of CZE-MS include the intact mass analysis of biopharmaceutical proteins11 and monoclonal antibodies22,68 where complete baseline separation is often achieved.”
From Recent trends of capillary electrophoresis‐mass spectrometry in proteomics research (2019) · cites this paper as [68]
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Self-citationOptimizing capillary electrophoresis for top‐down proteomics of 30–80 kDa proteins
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Ion Activation Methods for Peptides and Proteins
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- Authors
- Zhao, Y., Sun, L., Knierman, M. D., & Dovichi, N. J.
- Affiliations
- University of Notre Dame (US)
- Eli Lilly (United States) (US)
- Venue
- Talanta
- Publisher
- Elsevier BV
- Published
- 2016-02
- Volume
- 148
- Pages
- 529-533
- ISSN
- 0039-9140 (ISSN-L)
- OpenAlex
- W2133061982
- Cited by
- 49
- References
- 7
- Citations by year
- 2024: 4 · 2023: 4 · 2022: 1 · 2021: 10 · 2020: 2 · 2019: 6 · 2018: 5 · 2017: 6 · 2016: 10 · 2015: 1
- FWCI
- 3.65
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Talanta (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Eli Lilly and Company
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- capillary zone electrophoresis, top-down proteomics, antibody heavy and light chains, background electrolyte optimization, antibody mixtures
- MeSH terms
- Antibodies, Monoclonal, Time Factors, Electrophoresis, Capillary, Tandem Mass Spectrometry
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