Direct measurement of light and heavy antibody chains using ion mobility and middle-down mass spectrometry

Rafael D. Melani; Kristina Srzentić; Vincent R. Gerbasi; John P. McGee; Romain Huguet; Luca Fornelli; Neil L. Kelleher

doi:10.1080/19420862.2019.1668226

Direct measurement of light and heavy antibody chains using ion mobility and middle-down mass spectrometry

Rafael D. Melani^*, Kristina Srzentić, Vincent R. Gerbasi, John P. McGee, Romain Huguet, Luca Fornelli, Neil L. Kelleher

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

The analysis of monoclonal antibodies (mAbs) by a middle-down mass spectrometry (MS) approach is a growing field that attracts the attention of many researchers and biopharmaceutical companies. Usually, liquid fractionation techniques are used to separate mAbs polypeptides chains before MS analysis. Gas-phase fractionation techniques such as high-field asymmetric waveform ion mobility spectrometry (FAIMS) can replace liquid-based separations and reduce both analysis time and cost. Here, we present a rapid FAIMS tandem MS method capable of characterizing the polypeptide sequence of mAbs light and heavy chains in an unprecedented, easy, and fast fashion. This new method uses commercially available instruments and takes ~24 min, which is 40-60% faster than regular liquid chromatography-MS/MS analysis, to acquire fragmentation data using different dissociation methods.

Original language	English (US)
Pages (from-to)	1351-1357
Number of pages	7
Journal	mAbs
Volume	11
Issue number	8
DOIs	https://doi.org/10.1080/19420862.2019.1668226
State	Published - Nov 17 2019

Funding

This research was carried out in collaboration with the National Resource for Translational and Developmental Proteomics under Grant P41 GM108569 from the National Institute of General Medical Sciences (NLK) and supported by the Sherman Fairchild Foundation. LF would like to thank the University of Oklahoma, Department of Biology, for start-up funds.

Keywords

FAIMS
gas-phase fractionation
light and heavy chains
mAb
middle-down analysis

ASJC Scopus subject areas

Immunology and Allergy
Immunology

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10.1080/19420862.2019.1668226

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abstract = "The analysis of monoclonal antibodies (mAbs) by a middle-down mass spectrometry (MS) approach is a growing field that attracts the attention of many researchers and biopharmaceutical companies. Usually, liquid fractionation techniques are used to separate mAbs polypeptides chains before MS analysis. Gas-phase fractionation techniques such as high-field asymmetric waveform ion mobility spectrometry (FAIMS) can replace liquid-based separations and reduce both analysis time and cost. Here, we present a rapid FAIMS tandem MS method capable of characterizing the polypeptide sequence of mAbs light and heavy chains in an unprecedented, easy, and fast fashion. This new method uses commercially available instruments and takes ~24 min, which is 40-60% faster than regular liquid chromatography-MS/MS analysis, to acquire fragmentation data using different dissociation methods.",

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author = "Melani, {Rafael D.} and Kristina Srzenti{\'c} and Gerbasi, {Vincent R.} and McGee, {John P.} and Romain Huguet and Luca Fornelli and Kelleher, {Neil L.}",

note = "Funding Information: This research was carried out in collaboration with the National Resource for Translational and Developmental Proteomics under Grant P41 GM108569 from the National Institute of General Medical Sciences (NLK) and supported by the Sherman Fairchild Foundation. LF would like to thank the University of Oklahoma, Department of Biology, for start-up funds. Publisher Copyright: {\textcopyright} 2019, {\textcopyright} 2019 The Author(s). Published with license by Taylor & Francis Group, LLC.",

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doi = "10.1080/19420862.2019.1668226",

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AU - Srzentić, Kristina

AU - Gerbasi, Vincent R.

AU - McGee, John P.

AU - Huguet, Romain

AU - Fornelli, Luca

AU - Kelleher, Neil L.

N1 - Funding Information: This research was carried out in collaboration with the National Resource for Translational and Developmental Proteomics under Grant P41 GM108569 from the National Institute of General Medical Sciences (NLK) and supported by the Sherman Fairchild Foundation. LF would like to thank the University of Oklahoma, Department of Biology, for start-up funds. Publisher Copyright: © 2019, © 2019 The Author(s). Published with license by Taylor & Francis Group, LLC.

PY - 2019/11/17

Y1 - 2019/11/17

N2 - The analysis of monoclonal antibodies (mAbs) by a middle-down mass spectrometry (MS) approach is a growing field that attracts the attention of many researchers and biopharmaceutical companies. Usually, liquid fractionation techniques are used to separate mAbs polypeptides chains before MS analysis. Gas-phase fractionation techniques such as high-field asymmetric waveform ion mobility spectrometry (FAIMS) can replace liquid-based separations and reduce both analysis time and cost. Here, we present a rapid FAIMS tandem MS method capable of characterizing the polypeptide sequence of mAbs light and heavy chains in an unprecedented, easy, and fast fashion. This new method uses commercially available instruments and takes ~24 min, which is 40-60% faster than regular liquid chromatography-MS/MS analysis, to acquire fragmentation data using different dissociation methods.

AB - The analysis of monoclonal antibodies (mAbs) by a middle-down mass spectrometry (MS) approach is a growing field that attracts the attention of many researchers and biopharmaceutical companies. Usually, liquid fractionation techniques are used to separate mAbs polypeptides chains before MS analysis. Gas-phase fractionation techniques such as high-field asymmetric waveform ion mobility spectrometry (FAIMS) can replace liquid-based separations and reduce both analysis time and cost. Here, we present a rapid FAIMS tandem MS method capable of characterizing the polypeptide sequence of mAbs light and heavy chains in an unprecedented, easy, and fast fashion. This new method uses commercially available instruments and takes ~24 min, which is 40-60% faster than regular liquid chromatography-MS/MS analysis, to acquire fragmentation data using different dissociation methods.

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Direct measurement of light and heavy antibody chains using ion mobility and middle-down mass spectrometry

Abstract

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Keywords

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