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Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles
Extracellular vesicles (EVs) are emerging fundamental players in viral infections by shuttling viral components, mediating immune responses and likely the spread of the virus. However, the obstacles involved in purifying EVs and removing contaminating viral particles in a reliable and effective mann...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7754750/ https://www.ncbi.nlm.nih.gov/pubmed/33363711 http://dx.doi.org/10.1002/jev2.12040 |
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author | Jung, Stephanie Jacobs, Karolin Fiona Kirsten Shein, Mikhail Schütz, Anne Kathrin Mohr, Fabian Stadler, Herbert Stadler, Daniela Lucko, Aaron Michael Altstetter, Sebastian Maximilian Wilsch, Florian Deng, Li Protzer, Ulrike |
author_facet | Jung, Stephanie Jacobs, Karolin Fiona Kirsten Shein, Mikhail Schütz, Anne Kathrin Mohr, Fabian Stadler, Herbert Stadler, Daniela Lucko, Aaron Michael Altstetter, Sebastian Maximilian Wilsch, Florian Deng, Li Protzer, Ulrike |
author_sort | Jung, Stephanie |
collection | PubMed |
description | Extracellular vesicles (EVs) are emerging fundamental players in viral infections by shuttling viral components, mediating immune responses and likely the spread of the virus. However, the obstacles involved in purifying EVs and removing contaminating viral particles in a reliable and effective manner bottlenecks the full potential for the development of clinical and diagnostic treatment options targeting EV. Because of the similarities in size, density, membrane composition and mode of biogenesis of EVs and virions there are no standardized approaches for virus‐removal from EV preparations yet. Functional EV studies also require EV samples that are devoid of antibody contaminants. Consequently, the study of EVs in virology needs reliable and effective protocols to purify EVs and remove contaminating antibodies and viral particles. Here, we established a protocol for EV purification from hepatitis B virus (HBV)‐containing plasma by a combination of size‐exclusion chromatography and affinity‐based purification. After purification, EV samples were free of virus‐sized particles, HBV surface antigen, HBV core antigen, antibodies or infectious material. Viral genomic contamination was also decreased following purification. By using appropriate antibodies and size parameters, this protocol could potentially be applied to purification of EVs from other viral samples. In summary, we established a fast, reproducible and robust approach for the removal of HBV from EV preparations. Looking forward to the point of purifying EVs from clinical samples, this method should enable studies shedding light on the underlying mechanisms of EVs in viral infections and their diagnostic and prognostic potential. |
format | Online Article Text |
id | pubmed-7754750 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-77547502020-12-23 Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles Jung, Stephanie Jacobs, Karolin Fiona Kirsten Shein, Mikhail Schütz, Anne Kathrin Mohr, Fabian Stadler, Herbert Stadler, Daniela Lucko, Aaron Michael Altstetter, Sebastian Maximilian Wilsch, Florian Deng, Li Protzer, Ulrike J Extracell Vesicles Technical Reports Extracellular vesicles (EVs) are emerging fundamental players in viral infections by shuttling viral components, mediating immune responses and likely the spread of the virus. However, the obstacles involved in purifying EVs and removing contaminating viral particles in a reliable and effective manner bottlenecks the full potential for the development of clinical and diagnostic treatment options targeting EV. Because of the similarities in size, density, membrane composition and mode of biogenesis of EVs and virions there are no standardized approaches for virus‐removal from EV preparations yet. Functional EV studies also require EV samples that are devoid of antibody contaminants. Consequently, the study of EVs in virology needs reliable and effective protocols to purify EVs and remove contaminating antibodies and viral particles. Here, we established a protocol for EV purification from hepatitis B virus (HBV)‐containing plasma by a combination of size‐exclusion chromatography and affinity‐based purification. After purification, EV samples were free of virus‐sized particles, HBV surface antigen, HBV core antigen, antibodies or infectious material. Viral genomic contamination was also decreased following purification. By using appropriate antibodies and size parameters, this protocol could potentially be applied to purification of EVs from other viral samples. In summary, we established a fast, reproducible and robust approach for the removal of HBV from EV preparations. Looking forward to the point of purifying EVs from clinical samples, this method should enable studies shedding light on the underlying mechanisms of EVs in viral infections and their diagnostic and prognostic potential. John Wiley and Sons Inc. 2020-12-22 2020-12 /pmc/articles/PMC7754750/ /pubmed/33363711 http://dx.doi.org/10.1002/jev2.12040 Text en © 2020 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals LLC on behalf of International Society for Extracellular Vesicles This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Technical Reports Jung, Stephanie Jacobs, Karolin Fiona Kirsten Shein, Mikhail Schütz, Anne Kathrin Mohr, Fabian Stadler, Herbert Stadler, Daniela Lucko, Aaron Michael Altstetter, Sebastian Maximilian Wilsch, Florian Deng, Li Protzer, Ulrike Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles |
title | Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles |
title_full | Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles |
title_fullStr | Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles |
title_full_unstemmed | Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles |
title_short | Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles |
title_sort | efficient and reproducible depletion of hepatitis b virus from plasma derived extracellular vesicles |
topic | Technical Reports |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7754750/ https://www.ncbi.nlm.nih.gov/pubmed/33363711 http://dx.doi.org/10.1002/jev2.12040 |
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