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Systematic LC/MS/MS Investigations for the IND-Enabling Extended Characterization of Antibody–Drug Conjugate Modifications

We hypothesized that systematic liquid chromatography-tandem mass spectrometry investigations of an antibody–drug conjugate (ADC), its small and large molecular components, and surrogate small-molecule conjugates might comprise a simple and efficient approach for the extended characterization of ADC...

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Autores principales: Linz, Thomas, Yeo, Dominick, Hong, Qiuting, Zmolek, Wesley, McFarland, Jesse, Barfield, Robyn M., Haskins, William E., Rabuka, David
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6698958/
https://www.ncbi.nlm.nih.gov/pubmed/31544890
http://dx.doi.org/10.3390/antib7040040
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author Linz, Thomas
Yeo, Dominick
Hong, Qiuting
Zmolek, Wesley
McFarland, Jesse
Barfield, Robyn M.
Haskins, William E.
Rabuka, David
author_facet Linz, Thomas
Yeo, Dominick
Hong, Qiuting
Zmolek, Wesley
McFarland, Jesse
Barfield, Robyn M.
Haskins, William E.
Rabuka, David
author_sort Linz, Thomas
collection PubMed
description We hypothesized that systematic liquid chromatography-tandem mass spectrometry investigations of an antibody–drug conjugate (ADC), its small and large molecular components, and surrogate small-molecule conjugates might comprise a simple and efficient approach for the extended characterization of ADCs. Furthermore, we envisioned that results from this work might allow us to assign specific composition changes in the ADC based on monoisotopic mass shifts of conjugatable modifications as detected in the surrogate small-molecule conjugates. We tested our hypothesis with a case study using an aldehyde-tag-based ADC conjugated to a noncleavable linker bearing a maytansine payload. Nearly quantitative bioconversion from cysteine to formylglycine was observed in the monoclonal antibody, and bioorthogonal conjugation was detected only on the formylglycine residues in the ADC. Using our method, both conjugatable and nonconjugatable modifications were discovered in the linker/payload; however, only conjugatable modifications were observed on the ADC. Based on these results, we anticipate that our approach to systematic mass spectrometric investigations can be successfully applied to other ADCs and therapeutic bioconjugates for investigational new drug (IND)-enabling extended characterization.
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spelling pubmed-66989582019-09-05 Systematic LC/MS/MS Investigations for the IND-Enabling Extended Characterization of Antibody–Drug Conjugate Modifications Linz, Thomas Yeo, Dominick Hong, Qiuting Zmolek, Wesley McFarland, Jesse Barfield, Robyn M. Haskins, William E. Rabuka, David Antibodies (Basel) Article We hypothesized that systematic liquid chromatography-tandem mass spectrometry investigations of an antibody–drug conjugate (ADC), its small and large molecular components, and surrogate small-molecule conjugates might comprise a simple and efficient approach for the extended characterization of ADCs. Furthermore, we envisioned that results from this work might allow us to assign specific composition changes in the ADC based on monoisotopic mass shifts of conjugatable modifications as detected in the surrogate small-molecule conjugates. We tested our hypothesis with a case study using an aldehyde-tag-based ADC conjugated to a noncleavable linker bearing a maytansine payload. Nearly quantitative bioconversion from cysteine to formylglycine was observed in the monoclonal antibody, and bioorthogonal conjugation was detected only on the formylglycine residues in the ADC. Using our method, both conjugatable and nonconjugatable modifications were discovered in the linker/payload; however, only conjugatable modifications were observed on the ADC. Based on these results, we anticipate that our approach to systematic mass spectrometric investigations can be successfully applied to other ADCs and therapeutic bioconjugates for investigational new drug (IND)-enabling extended characterization. MDPI 2018-11-16 /pmc/articles/PMC6698958/ /pubmed/31544890 http://dx.doi.org/10.3390/antib7040040 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Linz, Thomas
Yeo, Dominick
Hong, Qiuting
Zmolek, Wesley
McFarland, Jesse
Barfield, Robyn M.
Haskins, William E.
Rabuka, David
Systematic LC/MS/MS Investigations for the IND-Enabling Extended Characterization of Antibody–Drug Conjugate Modifications
title Systematic LC/MS/MS Investigations for the IND-Enabling Extended Characterization of Antibody–Drug Conjugate Modifications
title_full Systematic LC/MS/MS Investigations for the IND-Enabling Extended Characterization of Antibody–Drug Conjugate Modifications
title_fullStr Systematic LC/MS/MS Investigations for the IND-Enabling Extended Characterization of Antibody–Drug Conjugate Modifications
title_full_unstemmed Systematic LC/MS/MS Investigations for the IND-Enabling Extended Characterization of Antibody–Drug Conjugate Modifications
title_short Systematic LC/MS/MS Investigations for the IND-Enabling Extended Characterization of Antibody–Drug Conjugate Modifications
title_sort systematic lc/ms/ms investigations for the ind-enabling extended characterization of antibody–drug conjugate modifications
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6698958/
https://www.ncbi.nlm.nih.gov/pubmed/31544890
http://dx.doi.org/10.3390/antib7040040
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