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  4. Drug-to-Antibody Ratio Estimation via Proteoform Peak Integration in the Analysis of Antibody-Oligonucleotide Conjugates with Orbitrap Fourier Transform Mass Spectrometry
 
research article

Drug-to-Antibody Ratio Estimation via Proteoform Peak Integration in the Analysis of Antibody-Oligonucleotide Conjugates with Orbitrap Fourier Transform Mass Spectrometry

Nagornov, Konstantin O.  
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Gasilova, Natalia  
•
Kozhinov, Anton N.  
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September 28, 2021
Analytical Chemistry

The therapeutic efficacy and pharmacokinetics of antibody-drug conjugates (ADCs) in general, and antibody-oligonucleotide conjugates (AOCs) in particular, depend on the drug-to-antibody ratio (DAR) distribution and average value. The DAR is considered a critical quality attribute, and information pertaining to it needs to be gathered during ADC/AOC development, production, and storage. However, because of the high structural complexity of ADC/AOC samples, particularly in the initial drug-development stages, the application of the current state-of-the-art mass spectrometric approaches can be limited for DAR analysis. Here, we demonstrate a novel approach for the analysis of complex ADC/AOC samples, following native size-exclusion chromatography Orbitrap Fourier transform mass spectrometry (FTMS). The approach is based on the integration of the proteoform-level mass spectral peaks in order to provide an estimate of the DAR distribution and its average value with less than 10% error. The peak integration is performed via a truncation of the Orbitrap's unreduced time-domain ion signals (transients) before mass spectra generation via FT processing. Transient recording and processing are undertaken using an external data acquisition system, FTMS Booster X2, coupled to a Q Exactive HF Orbitrap FTMS instrument. This approach has been applied to the analysis of whole and subunit-level trastuzumab conjugates with oligonucleotides. The obtained results indicate that ADC/AOC sample purification or simplification procedures, for example, deglycosylation, could be omitted or minimized prior to the DAR analysis, streamlining the drug-development process.

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Type
research article
DOI
10.1021/acs.analchem.1c02247
Web of Science ID

WOS:000703526400016

Author(s)
Nagornov, Konstantin O.  
Gasilova, Natalia  
Kozhinov, Anton N.  
Virta, Pasi
Holm, Patrik
Menin, Laure  
Nesatyy, Victor J.
Tsybin, Yury O.  
Date Issued

2021-09-28

Publisher

AMER CHEMICAL SOC

Published in
Analytical Chemistry
Volume

93

Issue

38

Start page

12930

End page

12937

Subjects

Chemistry, Analytical

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Chemistry

•

resolution

•

mobility

•

deconvolution

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strategies

•

ions

•

ms

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

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LEPA  
ISIC-GE  
Available on Infoscience
October 23, 2021
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/182479
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