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  4. Characterization of the Time-Domain Isotopic Beat Patterns of Monoclonal Antibodies in Fourier Transform Mass Spectrometry
 
research article

Characterization of the Time-Domain Isotopic Beat Patterns of Monoclonal Antibodies in Fourier Transform Mass Spectrometry

Nagornov, Konstantin O.
•
Kozhinov, Anton N.
•
Gasilova, Natalia  
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May 31, 2022
Journal Of The American Society For Mass Spectrometry

The time-domain transients in the Fourier transform mass spectrometry (FTMS) analysis of monoclonal antibodies (mAbs) are known to exhibit characteristic isotopic beat patterns. These patterns are defined by the isotopic distributions of all gaseous mAb ions present in the FTMS mass analyzer, originating from single or multiple charge states, and from single or multiple proteoforms. For an isolated charge state of a single proteofonn, the mAb isotopic beat pattern resembles narrow splashes of signal amplitude (beats), spaced periodically in the time-domain transient, with broad (often exceeding 1 s) "valleys" between them. Here, we reinforce the importance of isotopic beat patterns for the accurate interpretation and presentation of FTMS data in the analysis of mAbs and other large biopolymers. An updated, mAb-grade version of the transient-mediated FTMS data simulation and visualization tool, FTMS Simulator is introduced and benchmarked. We then apply this tool to evaluate the charge-state dependent characteristics of isotopic beats in mAbs analyses with modern models of Orbitrap and ion cydotron resonance (ICR) FTMS instruments, including detection of higher-order harmonics. We demonstrate the impact of the isotopic beat patterns on the analytical characteristics of the resulting mass spectra of individual and overlapping mAb proteoforms. The results reported here detail highly nonlinear dependences of resolution and signalto-noise ratio on the time-domain transient period, absorption or magnitude mode spectra representation, and apodization functions. The provided description and the demonstrated ability to routinely conduct accurate simulations of FTMS data for large biopolymers should aid the end-users of Orbitrap and ICR FTMS instruments in the analysis of mAbs and other biopolymers, including viruses.

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Type
research article
DOI
10.1021/jasms.1c00336
Web of Science ID

WOS:000821173500001

Author(s)
Nagornov, Konstantin O.
Kozhinov, Anton N.
Gasilova, Natalia  
Menin, Laure  
Tsybin, Yury O.
Date Issued

2022-05-31

Published in
Journal Of The American Society For Mass Spectrometry
Subjects

Biochemical Research Methods

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Chemistry, Analytical

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Chemistry, Physical

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Spectroscopy

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Biochemistry & Molecular Biology

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Chemistry

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Spectroscopy

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time-domain transient

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fourier transform mass spectrometry

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ftms

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orbitrap

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ion cyclotron resonance

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icr

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electron-transfer dissociation

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top-down analysis

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structural-analysis

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resolution

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protein

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acquisition

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performance

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spectra

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
ISIC-MSEAP  
Available on Infoscience
August 1, 2022
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/189551
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