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Use of a charge reducing agent to enable intact mass analysis of cysteine-linked antibody-drug-conjugates by native mass spectrometry

Antibody-drug-conjugates (ADC) are a growing class of anticancer biopharmaceuticals. Conjugation of cysteine linked ADCs, requires initial reduction of mAb inter-chain disulfide bonds, as the drugs are attached via thiol chemistry. This results in the active mAb moiety being transformed from a coval...

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Autores principales: Pacholarz, Kamila J., Barran, Perdita E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5988552/
https://www.ncbi.nlm.nih.gov/pubmed/29900109
http://dx.doi.org/10.1016/j.euprot.2016.02.004
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author Pacholarz, Kamila J.
Barran, Perdita E.
author_facet Pacholarz, Kamila J.
Barran, Perdita E.
author_sort Pacholarz, Kamila J.
collection PubMed
description Antibody-drug-conjugates (ADC) are a growing class of anticancer biopharmaceuticals. Conjugation of cysteine linked ADCs, requires initial reduction of mAb inter-chain disulfide bonds, as the drugs are attached via thiol chemistry. This results in the active mAb moiety being transformed from a covalently linked tetramer to non-covalently linked complexes, which hinders precise determination of drug load with LC–MS. Here, we show how the addition of the charge reducing agent triethylammonium acetate (TEAA) preserves the intact mAb structure, is well suited to the study of cysteine linked conjugates and facilitates easy drug load determination by direct infusion native MS.
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spelling pubmed-59885522018-06-13 Use of a charge reducing agent to enable intact mass analysis of cysteine-linked antibody-drug-conjugates by native mass spectrometry Pacholarz, Kamila J. Barran, Perdita E. EuPA Open Proteom Special Section: Proceedings of the 9th Annual EuPA Congress “Proteomics - Back to the Future” (June 23 - 28, 2015, Milano, Italy) Antibody-drug-conjugates (ADC) are a growing class of anticancer biopharmaceuticals. Conjugation of cysteine linked ADCs, requires initial reduction of mAb inter-chain disulfide bonds, as the drugs are attached via thiol chemistry. This results in the active mAb moiety being transformed from a covalently linked tetramer to non-covalently linked complexes, which hinders precise determination of drug load with LC–MS. Here, we show how the addition of the charge reducing agent triethylammonium acetate (TEAA) preserves the intact mAb structure, is well suited to the study of cysteine linked conjugates and facilitates easy drug load determination by direct infusion native MS. Elsevier 2016-03-04 /pmc/articles/PMC5988552/ /pubmed/29900109 http://dx.doi.org/10.1016/j.euprot.2016.02.004 Text en © 2016 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Special Section: Proceedings of the 9th Annual EuPA Congress “Proteomics - Back to the Future” (June 23 - 28, 2015, Milano, Italy)
Pacholarz, Kamila J.
Barran, Perdita E.
Use of a charge reducing agent to enable intact mass analysis of cysteine-linked antibody-drug-conjugates by native mass spectrometry
title Use of a charge reducing agent to enable intact mass analysis of cysteine-linked antibody-drug-conjugates by native mass spectrometry
title_full Use of a charge reducing agent to enable intact mass analysis of cysteine-linked antibody-drug-conjugates by native mass spectrometry
title_fullStr Use of a charge reducing agent to enable intact mass analysis of cysteine-linked antibody-drug-conjugates by native mass spectrometry
title_full_unstemmed Use of a charge reducing agent to enable intact mass analysis of cysteine-linked antibody-drug-conjugates by native mass spectrometry
title_short Use of a charge reducing agent to enable intact mass analysis of cysteine-linked antibody-drug-conjugates by native mass spectrometry
title_sort use of a charge reducing agent to enable intact mass analysis of cysteine-linked antibody-drug-conjugates by native mass spectrometry
topic Special Section: Proceedings of the 9th Annual EuPA Congress “Proteomics - Back to the Future” (June 23 - 28, 2015, Milano, Italy)
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5988552/
https://www.ncbi.nlm.nih.gov/pubmed/29900109
http://dx.doi.org/10.1016/j.euprot.2016.02.004
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