A Native Mass Spectrometry-Based Assay for Rapid Assessment of the Empty:Full Capsid Ratio in Adeno-Associated Virus Gene Therapy Products

[Image: see text] Adeno-associated virus (AAV)-based gene therapy is a rapidly developing field, requiring analytical methods for detailed product characterization. One important quality attribute of AAV products that requires monitoring is the amount of residual empty capsids following downstream p...

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Autores principales: Strasser, Lisa, Morgan, Tomos E., Guapo, Felipe, Füssl, Florian, Forsey, Daniel, Anderson, Ian, Bones, Jonathan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2021
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8482367/
https://www.ncbi.nlm.nih.gov/pubmed/34519199
http://dx.doi.org/10.1021/acs.analchem.1c02828
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author Strasser, Lisa
Morgan, Tomos E.
Guapo, Felipe
Füssl, Florian
Forsey, Daniel
Anderson, Ian
Bones, Jonathan
author_facet Strasser, Lisa
Morgan, Tomos E.
Guapo, Felipe
Füssl, Florian
Forsey, Daniel
Anderson, Ian
Bones, Jonathan
author_sort Strasser, Lisa
collection PubMed
description [Image: see text] Adeno-associated virus (AAV)-based gene therapy is a rapidly developing field, requiring analytical methods for detailed product characterization. One important quality attribute of AAV products that requires monitoring is the amount of residual empty capsids following downstream processing. Traditionally, empty and full particles are quantified via analytical ultracentrifugation as well as anion exchange chromatography using ultraviolet or fluorescence detection. Here, we present a native mass spectrometry-based approach to assess the ratio of empty to full AAV-capsids without the need for excessive sample preparation. We report the rapid determination of the relative amount of empty capsids in AAV5 and AAV8 samples. The results correlate well with more conventional analysis strategies, demonstrating the potential of native mass spectrometry for the characterization of viral particles.
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spelling pubmed-84823672021-09-30 A Native Mass Spectrometry-Based Assay for Rapid Assessment of the Empty:Full Capsid Ratio in Adeno-Associated Virus Gene Therapy Products Strasser, Lisa Morgan, Tomos E. Guapo, Felipe Füssl, Florian Forsey, Daniel Anderson, Ian Bones, Jonathan Anal Chem [Image: see text] Adeno-associated virus (AAV)-based gene therapy is a rapidly developing field, requiring analytical methods for detailed product characterization. One important quality attribute of AAV products that requires monitoring is the amount of residual empty capsids following downstream processing. Traditionally, empty and full particles are quantified via analytical ultracentrifugation as well as anion exchange chromatography using ultraviolet or fluorescence detection. Here, we present a native mass spectrometry-based approach to assess the ratio of empty to full AAV-capsids without the need for excessive sample preparation. We report the rapid determination of the relative amount of empty capsids in AAV5 and AAV8 samples. The results correlate well with more conventional analysis strategies, demonstrating the potential of native mass spectrometry for the characterization of viral particles. American Chemical Society 2021-09-14 2021-09-28 /pmc/articles/PMC8482367/ /pubmed/34519199 http://dx.doi.org/10.1021/acs.analchem.1c02828 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Strasser, Lisa
Morgan, Tomos E.
Guapo, Felipe
Füssl, Florian
Forsey, Daniel
Anderson, Ian
Bones, Jonathan
A Native Mass Spectrometry-Based Assay for Rapid Assessment of the Empty:Full Capsid Ratio in Adeno-Associated Virus Gene Therapy Products
title A Native Mass Spectrometry-Based Assay for Rapid Assessment of the Empty:Full Capsid Ratio in Adeno-Associated Virus Gene Therapy Products
title_full A Native Mass Spectrometry-Based Assay for Rapid Assessment of the Empty:Full Capsid Ratio in Adeno-Associated Virus Gene Therapy Products
title_fullStr A Native Mass Spectrometry-Based Assay for Rapid Assessment of the Empty:Full Capsid Ratio in Adeno-Associated Virus Gene Therapy Products
title_full_unstemmed A Native Mass Spectrometry-Based Assay for Rapid Assessment of the Empty:Full Capsid Ratio in Adeno-Associated Virus Gene Therapy Products
title_short A Native Mass Spectrometry-Based Assay for Rapid Assessment of the Empty:Full Capsid Ratio in Adeno-Associated Virus Gene Therapy Products
title_sort native mass spectrometry-based assay for rapid assessment of the empty:full capsid ratio in adeno-associated virus gene therapy products
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8482367/
https://www.ncbi.nlm.nih.gov/pubmed/34519199
http://dx.doi.org/10.1021/acs.analchem.1c02828
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