Cited by 67Mass Spectrometry Techniques and ApplicationsAdvanced Proteomics Techniques and ApplicationsMetabolomics and Mass Spectrometry Studies

Unit Mass Baseline Resolution for an Intact 148 kDa Therapeutic Monoclonal Antibody by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

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Valeja, Santosh G., Kaiser, Nathan K., Xian, Feng, Hendrickson, Christopher L., Rouse, Jason C., Marshall, Alan G.·2011·Analytical Chemistry

Abstract

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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Crossref · 10.1021/ac202429c

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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.”

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  1. [15]Kelleher NL Anal. Chem 2004 76 197A 203A 15190879
  2. [49]Marshall AG Hendrickson CL Rapid Commun. Mass Spectrom 2001 15 232 235 10421907

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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
Published
2011-10-20
Volume
83
Issue
22
Pages
8391-8395
ISSN
  • 0003-2700 (ISSN-L)
  • 1520-6882
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
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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