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High-throughput screening of antibody variants for chemical stability: identification of deamidation-resistant mutants

Developability assessment of therapeutic antibody candidates assists drug discovery by enabling early identification of undesirable instabilities. Rapid chemical stability screening of antibody variants can accelerate the identification of potential solutions. We describe here the development of a h...

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Autores principales: DiCara, Danielle M., Andersen, Nisana, Chan, Ruby, Ernst, James A., Ayalon, Gai, Lazar, Greg A., Agard, Nicholas J., Hilderbrand, Amy, Hötzel, Isidro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6204805/
https://www.ncbi.nlm.nih.gov/pubmed/30130444
http://dx.doi.org/10.1080/19420862.2018.1504726
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author DiCara, Danielle M.
Andersen, Nisana
Chan, Ruby
Ernst, James A.
Ayalon, Gai
Lazar, Greg A.
Agard, Nicholas J.
Hilderbrand, Amy
Hötzel, Isidro
author_facet DiCara, Danielle M.
Andersen, Nisana
Chan, Ruby
Ernst, James A.
Ayalon, Gai
Lazar, Greg A.
Agard, Nicholas J.
Hilderbrand, Amy
Hötzel, Isidro
author_sort DiCara, Danielle M.
collection PubMed
description Developability assessment of therapeutic antibody candidates assists drug discovery by enabling early identification of undesirable instabilities. Rapid chemical stability screening of antibody variants can accelerate the identification of potential solutions. We describe here the development of a high-throughput assay to characterize asparagine deamidation. We applied the assay to identify a mutation that unexpectedly stabilizes a critical asparagine. Ninety antibody variants were incubated under thermal stress in order to induce deamidation and screened for both affinity and total binding capacity. Surprisingly, a mutation five residues downstream from the unstable asparagine greatly reduced deamidation. Detailed assessment by LC-MS analysis confirmed the predicted improvement. This work describes both a high-throughput method for antibody stability screening during the early stages of antibody discovery and highlights the value of broad searches of antibody sequence space.
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spelling pubmed-62048052018-10-30 High-throughput screening of antibody variants for chemical stability: identification of deamidation-resistant mutants DiCara, Danielle M. Andersen, Nisana Chan, Ruby Ernst, James A. Ayalon, Gai Lazar, Greg A. Agard, Nicholas J. Hilderbrand, Amy Hötzel, Isidro MAbs Report Developability assessment of therapeutic antibody candidates assists drug discovery by enabling early identification of undesirable instabilities. Rapid chemical stability screening of antibody variants can accelerate the identification of potential solutions. We describe here the development of a high-throughput assay to characterize asparagine deamidation. We applied the assay to identify a mutation that unexpectedly stabilizes a critical asparagine. Ninety antibody variants were incubated under thermal stress in order to induce deamidation and screened for both affinity and total binding capacity. Surprisingly, a mutation five residues downstream from the unstable asparagine greatly reduced deamidation. Detailed assessment by LC-MS analysis confirmed the predicted improvement. This work describes both a high-throughput method for antibody stability screening during the early stages of antibody discovery and highlights the value of broad searches of antibody sequence space. Taylor & Francis 2018-09-11 /pmc/articles/PMC6204805/ /pubmed/30130444 http://dx.doi.org/10.1080/19420862.2018.1504726 Text en © 2018 The Author(s). Published by Taylor & Francis. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.
spellingShingle Report
DiCara, Danielle M.
Andersen, Nisana
Chan, Ruby
Ernst, James A.
Ayalon, Gai
Lazar, Greg A.
Agard, Nicholas J.
Hilderbrand, Amy
Hötzel, Isidro
High-throughput screening of antibody variants for chemical stability: identification of deamidation-resistant mutants
title High-throughput screening of antibody variants for chemical stability: identification of deamidation-resistant mutants
title_full High-throughput screening of antibody variants for chemical stability: identification of deamidation-resistant mutants
title_fullStr High-throughput screening of antibody variants for chemical stability: identification of deamidation-resistant mutants
title_full_unstemmed High-throughput screening of antibody variants for chemical stability: identification of deamidation-resistant mutants
title_short High-throughput screening of antibody variants for chemical stability: identification of deamidation-resistant mutants
title_sort high-throughput screening of antibody variants for chemical stability: identification of deamidation-resistant mutants
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6204805/
https://www.ncbi.nlm.nih.gov/pubmed/30130444
http://dx.doi.org/10.1080/19420862.2018.1504726
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