Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles
| Autor: | Aaron Michael Lucko, Herbert Stadler, Anne K Schütz, Daniela Stadler, Florian Wilsch, Mikhail Shein, Karolin Fiona Kirsten Jacobs, Sebastian Maximilian Altstetter, Stephanie Jung, Fabian Mohr, Ulrike Protzer, Li Deng |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
virus removal
0301 basic medicine Hepatitis B virus Histology Technical Reports viruses Ev Purification Affinity‐based Purification Extracellular Vesicles Hepatitis B Virus Size‐exclusion Chromatography Virus Removal EV purification medicine.disease_cause Extracellular vesicles Chromatography Affinity Virus Plasma 03 medical and health sciences Technical Report 0302 clinical medicine Immune system Antigen medicine Humans affinity‐based purification biology Chemistry Plasma derived Treatment options Cell Biology Hepatitis B Virology 030104 developmental biology 030220 oncology & carcinogenesis Chromatography Gel biology.protein Antibody size‐exclusion chromatography |
| Zdroj: | J. Extra. Vesicles 10:e12040 (2020) Journal of Extracellular Vesicles |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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