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Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles.
J. Extra. Vesicles 10:e12040 (2020)
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.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Ev Purification ; Affinity‐based Purification ; Extracellular Vesicles ; Hepatitis B Virus ; Size‐exclusion Chromatography ; Virus Removal
ISSN (print) / ISBN
2001-3078
e-ISSN
2001-3078
Quellenangaben
Volume: 10,
Issue: 2,
Article Number: e12040
Publisher
Taylor & Francis
Publishing Place
[S.l.]
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Institute of Virology (VIRO)
Grants
German Research Association (DFG)