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Size Exclusion Chromatography–Mass Photometry: A New Method for Adeno-Associated Virus Product Characterization
Over the past decade, adeno-associated viruses (AAVs) have attained significant prominence in gene therapy and genome editing applications, necessitating the development of robust and precise methodologies to ensure the quality and purity of AAV products. Existing AAV characterization techniques hav...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10528216/ https://www.ncbi.nlm.nih.gov/pubmed/37759487 http://dx.doi.org/10.3390/cells12182264 |
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author | Wu, Di Zhao, Xiaonan Jimenez, Diego Antonio Piszczek, Grzegorz |
author_facet | Wu, Di Zhao, Xiaonan Jimenez, Diego Antonio Piszczek, Grzegorz |
author_sort | Wu, Di |
collection | PubMed |
description | Over the past decade, adeno-associated viruses (AAVs) have attained significant prominence in gene therapy and genome editing applications, necessitating the development of robust and precise methodologies to ensure the quality and purity of AAV products. Existing AAV characterization techniques have proven effective for the analysis of pure and homogeneous AAV samples. However, there is still a demand for a rapid and low-sample-consumption method suitable for the characterization of lower purity or heterogeneous AAV samples commonly encountered in AAV products. Addressing this challenge, we propose the SEC-MP method, which combines size exclusion chromatography (SEC) with mass photometry (MP). In this novel approach, SEC effectively separates monomeric AAV particles from impurities, while the UV detector determines the virus particle concentration. MP complements this process by estimating the fraction of fully packaged AAVs in the total population of AAV particles. This combined methodology enables accurate determination of the titer of effective, fully packaged AAVs in samples containing aggregates, incorrectly packaged AAVs with incomplete genomes, protein or DNA fragments, and other impurities. Our experimental results demonstrate that SEC-MP provides valuable guidance for sample quality control and subsequent applications in the field of AAV research. |
format | Online Article Text |
id | pubmed-10528216 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-105282162023-09-28 Size Exclusion Chromatography–Mass Photometry: A New Method for Adeno-Associated Virus Product Characterization Wu, Di Zhao, Xiaonan Jimenez, Diego Antonio Piszczek, Grzegorz Cells Article Over the past decade, adeno-associated viruses (AAVs) have attained significant prominence in gene therapy and genome editing applications, necessitating the development of robust and precise methodologies to ensure the quality and purity of AAV products. Existing AAV characterization techniques have proven effective for the analysis of pure and homogeneous AAV samples. However, there is still a demand for a rapid and low-sample-consumption method suitable for the characterization of lower purity or heterogeneous AAV samples commonly encountered in AAV products. Addressing this challenge, we propose the SEC-MP method, which combines size exclusion chromatography (SEC) with mass photometry (MP). In this novel approach, SEC effectively separates monomeric AAV particles from impurities, while the UV detector determines the virus particle concentration. MP complements this process by estimating the fraction of fully packaged AAVs in the total population of AAV particles. This combined methodology enables accurate determination of the titer of effective, fully packaged AAVs in samples containing aggregates, incorrectly packaged AAVs with incomplete genomes, protein or DNA fragments, and other impurities. Our experimental results demonstrate that SEC-MP provides valuable guidance for sample quality control and subsequent applications in the field of AAV research. MDPI 2023-09-13 /pmc/articles/PMC10528216/ /pubmed/37759487 http://dx.doi.org/10.3390/cells12182264 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Wu, Di Zhao, Xiaonan Jimenez, Diego Antonio Piszczek, Grzegorz Size Exclusion Chromatography–Mass Photometry: A New Method for Adeno-Associated Virus Product Characterization |
title | Size Exclusion Chromatography–Mass Photometry: A New Method for Adeno-Associated Virus Product Characterization |
title_full | Size Exclusion Chromatography–Mass Photometry: A New Method for Adeno-Associated Virus Product Characterization |
title_fullStr | Size Exclusion Chromatography–Mass Photometry: A New Method for Adeno-Associated Virus Product Characterization |
title_full_unstemmed | Size Exclusion Chromatography–Mass Photometry: A New Method for Adeno-Associated Virus Product Characterization |
title_short | Size Exclusion Chromatography–Mass Photometry: A New Method for Adeno-Associated Virus Product Characterization |
title_sort | size exclusion chromatography–mass photometry: a new method for adeno-associated virus product characterization |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10528216/ https://www.ncbi.nlm.nih.gov/pubmed/37759487 http://dx.doi.org/10.3390/cells12182264 |
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