Rapid and Robust Continuous Purification of High-Titer Hepatitis B Virus for In Vitro and In Vivo Applications.

HBV for in vitro and in vivo assays HBV high-titer virus stocks heparin-affinity chromatography hepatitis B virus sucrose-gradient ultracentrifugation virus purification

Journal

Viruses
ISSN: 1999-4915
Titre abrégé: Viruses
Pays: Switzerland
ID NLM: 101509722

Informations de publication

Date de publication:
30 07 2021
Historique:
received: 09 06 2021
revised: 06 07 2021
accepted: 21 07 2021
entrez: 28 8 2021
pubmed: 29 8 2021
medline: 3 2 2022
Statut: epublish

Résumé

Available treatments for hepatitis B can control the virus but are rarely curative. This led to a global initiative to design new curative therapies for the 257 million patients affected. Discovery and development of these new therapies is contingent upon functional in vitro and in vivo hepatitis B virus (HBV) infection models. However, low titer and impurity of conventional HBV stocks reduce significance of in vitro infections and moreover limit challenge doses in current in vivo models. Therefore, there is a critical need for a robust, simple and reproducible protocol to generate high-purity and high-titer infectious HBV stocks. Here, we outline a three-step protocol for continuous production of high-quality HBV stocks from supernatants of HBV-replicating cell lines. This purification process takes less than 6 h, yields to high-titer stocks (up to 1 × 10

Identifiants

pubmed: 34452368
pii: v13081503
doi: 10.3390/v13081503
pmc: PMC8402639
pii:
doi:

Substances chimiques

DNA, Viral 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

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Auteurs

Jochen M Wettengel (JM)

Institute of Virology, Technische Universität München/Helmholtz Zentrum München, 81675 Munich, Germany.
Vaccine & Gene Therapy Institute, Oregon Health & Science University, Beaverton, OR 97006, USA.

Bianca Linden (B)

Institute of Virology, Technische Universität München/Helmholtz Zentrum München, 81675 Munich, Germany.

Knud Esser (K)

Institute of Virology, Technische Universität München/Helmholtz Zentrum München, 81675 Munich, Germany.

Michael Laue (M)

Advanced Light and Electron Microscopy (ZBS 4), Robert Koch Institute, 13353 Berlin, Germany.

Benjamin J Burwitz (BJ)

Vaccine & Gene Therapy Institute, Oregon Health & Science University, Beaverton, OR 97006, USA.

Ulrike Protzer (U)

Institute of Virology, Technische Universität München/Helmholtz Zentrum München, 81675 Munich, Germany.

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Classifications MeSH