Unit Mass Baseline Resolution for an Intact 148 kDa Therapeutic Monoclonal Antibody by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry
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Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) provides the highest mass resolving power and mass measurement accuracy for unambiguous identification of biomolecules. Previously, the highest-mass protein for which FTICR unit mass resolution had been obtained was 115 kDa at 7 T. Here, we present baseline resolution for an intact 147.7 kDa monoclonal antibody (mAb), by prior dissociation of noncovalent adducts, optimization of detected total ion number, and optimization of ICR cell parameters to minimize space charge shifts, peak coalescence, and destructive ion cloud Coulombic interactions. The resultant long ICR transient lifetime (as high as 20 s) results in magnitude-mode mass resolving power of ~420,000 at m/z 2,593 for the 57+ charge state (the highest mass for which baseline unit mass resolution has been achieved), auguring for future characterization of even larger intact proteins and protein complexes by FTICR MS. We also demonstrate up to 80% higher resolving power by phase correction to yield an absorption-mode mass spectrum.
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FTICR mass spectrometers are traditionally the instrument of choice for top-down because they resolve very large species, but slower scan speed can limit LC-MS/MS and global protein characterization.
“Traditionally, Fourier-transform ion cyclotron resonance (FTICR) mass spectrometers are the instrument of choice for top-down because of its capabilities to resolve very large species [96]; however, the slower scan speed can limit its capabilities for effective LC-MS/MS analysis and global protein characterization.”
From Top-down proteomics: challenges, innovations, and applications in basic and clinical research (2020) · cites this paper as [96]
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- Authors
- Valeja, S. G., Kaiser, N. K., Xian, F., Hendrickson, C. L., Rouse, J. C., & Marshall, A. G.
- Affiliations
- Florida State University (US)
- Pfizer (United States) (US)
- Corresponding author
- Hendrickson, Christopher L.
- Marshall, Alan G.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2011-10-20
- Volume
- 83
- Issue
- 22
- Pages
- 8391-8395
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- PMC
- PMC3215840
- OpenAlex
- W66210199
- Cited by
- 67
- References
- 72
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 1 · 2023: 4 · 2022: 2 · 2020: 8 · 2019: 2 · 2018: 3 · 2017: 1 · 2016: 5 · 2015: 5 · 2014: 7 · 2013: 15 · 2012: 11
- FWCI
- 4.78
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Analytical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (5r01gm031683-17)
- National Institute of General Medical Sciences (R01 GM031683, 1R01 GM78359, R01GM078359)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Keywords
- monoclonal antibodies, FT-ICR mass spectrometry, phase correction
- MeSH terms
- Antibodies, Monoclonal, Fourier Analysis, Molecular Weight, Recombinant Proteins, Mass Spectrometry
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