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Monoclonal antibody stability can be usefully monitored using the excitation-energy-dependent fluorescence edge-shift
Among the major challenges in the development of biopharmaceuticals are structural heterogeneity and aggregation. The development of a successful therapeutic monoclonal antibody (mAb) requires both a highly active and also stable molecule. Whilst a range of experimental (biophysical) approaches exis...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7527260/ https://www.ncbi.nlm.nih.gov/pubmed/32869839 http://dx.doi.org/10.1042/BCJ20200580 |
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author | Knight, Michael J. Woolley, Rachel E. Kwok, Anthony Parsons, Stuart Jones, Hannah B. L. Gulácsy, Christina E. Phaal, Polly Kassaar, Omar Dawkins, Kieran Rodriguez, Elizabeth Marques, Andreia Bowsher, Leo Wells, Stephen A. Watts, Andrew vandenElsen, Jean M. H. Turner, Alison O'Hara, John Pudney, Christopher R. |
author_facet | Knight, Michael J. Woolley, Rachel E. Kwok, Anthony Parsons, Stuart Jones, Hannah B. L. Gulácsy, Christina E. Phaal, Polly Kassaar, Omar Dawkins, Kieran Rodriguez, Elizabeth Marques, Andreia Bowsher, Leo Wells, Stephen A. Watts, Andrew vandenElsen, Jean M. H. Turner, Alison O'Hara, John Pudney, Christopher R. |
author_sort | Knight, Michael J. |
collection | PubMed |
description | Among the major challenges in the development of biopharmaceuticals are structural heterogeneity and aggregation. The development of a successful therapeutic monoclonal antibody (mAb) requires both a highly active and also stable molecule. Whilst a range of experimental (biophysical) approaches exist to track changes in stability of proteins, routine prediction of stability remains challenging. The fluorescence red edge excitation shift (REES) phenomenon is sensitive to a range of changes in protein structure. Based on recent work, we have found that quantifying the REES effect is extremely sensitive to changes in protein conformational state and dynamics. Given the extreme sensitivity, potentially this tool could provide a ‘fingerprint’ of the structure and stability of a protein. Such a tool would be useful in the discovery and development of biopharamceuticals and so we have explored our hypothesis with a panel of therapeutic mAbs. We demonstrate that the quantified REES data show remarkable sensitivity, being able to discern between structurally identical antibodies and showing sensitivity to unfolding and aggregation. The approach works across a broad concentration range (µg–mg/ml) and is highly consistent. We show that the approach can be applied alongside traditional characterisation testing within the context of a forced degradation study (FDS). Most importantly, we demonstrate the approach is able to predict the stability of mAbs both in the short (hours), medium (days) and long-term (months). The quantified REES data will find immediate use in the biopharmaceutical industry in quality assurance, formulation and development. The approach benefits from low technical complexity, is rapid and uses instrumentation which exists in most biochemistry laboratories without modification. |
format | Online Article Text |
id | pubmed-7527260 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-75272602020-10-06 Monoclonal antibody stability can be usefully monitored using the excitation-energy-dependent fluorescence edge-shift Knight, Michael J. Woolley, Rachel E. Kwok, Anthony Parsons, Stuart Jones, Hannah B. L. Gulácsy, Christina E. Phaal, Polly Kassaar, Omar Dawkins, Kieran Rodriguez, Elizabeth Marques, Andreia Bowsher, Leo Wells, Stephen A. Watts, Andrew vandenElsen, Jean M. H. Turner, Alison O'Hara, John Pudney, Christopher R. Biochem J Biophysics Among the major challenges in the development of biopharmaceuticals are structural heterogeneity and aggregation. The development of a successful therapeutic monoclonal antibody (mAb) requires both a highly active and also stable molecule. Whilst a range of experimental (biophysical) approaches exist to track changes in stability of proteins, routine prediction of stability remains challenging. The fluorescence red edge excitation shift (REES) phenomenon is sensitive to a range of changes in protein structure. Based on recent work, we have found that quantifying the REES effect is extremely sensitive to changes in protein conformational state and dynamics. Given the extreme sensitivity, potentially this tool could provide a ‘fingerprint’ of the structure and stability of a protein. Such a tool would be useful in the discovery and development of biopharamceuticals and so we have explored our hypothesis with a panel of therapeutic mAbs. We demonstrate that the quantified REES data show remarkable sensitivity, being able to discern between structurally identical antibodies and showing sensitivity to unfolding and aggregation. The approach works across a broad concentration range (µg–mg/ml) and is highly consistent. We show that the approach can be applied alongside traditional characterisation testing within the context of a forced degradation study (FDS). Most importantly, we demonstrate the approach is able to predict the stability of mAbs both in the short (hours), medium (days) and long-term (months). The quantified REES data will find immediate use in the biopharmaceutical industry in quality assurance, formulation and development. The approach benefits from low technical complexity, is rapid and uses instrumentation which exists in most biochemistry laboratories without modification. Portland Press Ltd. 2020-09-30 2020-09-28 /pmc/articles/PMC7527260/ /pubmed/32869839 http://dx.doi.org/10.1042/BCJ20200580 Text en © 2020 The Author(s) https://creativecommons.org/licenses/by/4.0/ This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) . Open access for this article was enabled by the participation of University of Bath in an all-inclusive Read & Publish pilot with Portland Press and the Biochemical Society under a transformative agreement with JISC. |
spellingShingle | Biophysics Knight, Michael J. Woolley, Rachel E. Kwok, Anthony Parsons, Stuart Jones, Hannah B. L. Gulácsy, Christina E. Phaal, Polly Kassaar, Omar Dawkins, Kieran Rodriguez, Elizabeth Marques, Andreia Bowsher, Leo Wells, Stephen A. Watts, Andrew vandenElsen, Jean M. H. Turner, Alison O'Hara, John Pudney, Christopher R. Monoclonal antibody stability can be usefully monitored using the excitation-energy-dependent fluorescence edge-shift |
title | Monoclonal antibody stability can be usefully monitored using the excitation-energy-dependent fluorescence edge-shift |
title_full | Monoclonal antibody stability can be usefully monitored using the excitation-energy-dependent fluorescence edge-shift |
title_fullStr | Monoclonal antibody stability can be usefully monitored using the excitation-energy-dependent fluorescence edge-shift |
title_full_unstemmed | Monoclonal antibody stability can be usefully monitored using the excitation-energy-dependent fluorescence edge-shift |
title_short | Monoclonal antibody stability can be usefully monitored using the excitation-energy-dependent fluorescence edge-shift |
title_sort | monoclonal antibody stability can be usefully monitored using the excitation-energy-dependent fluorescence edge-shift |
topic | Biophysics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7527260/ https://www.ncbi.nlm.nih.gov/pubmed/32869839 http://dx.doi.org/10.1042/BCJ20200580 |
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