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

Wettengel, Jochen M.; Linden, Bianca; Esser, Knud; Laue, Michael; Burwitz, Benjamin J.; Protzer, Ulrike

doi:10.3390/v13081503

Open AccessProtocol

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

by

Jochen M. Wettengel

^1,2

,

Bianca Linden

¹

,

Knud Esser

¹,

Michael Laue

³

,

Benjamin J. Burwitz

²

and

Ulrike Protzer

^1,*

¹

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

²

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

³

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

^*

Author to whom correspondence should be addressed.

Viruses 2021, 13(8), 1503; https://doi.org/10.3390/v13081503

Submission received: 9 June 2021 / Revised: 6 July 2021 / Accepted: 21 July 2021 / Published: 30 July 2021

(This article belongs to the Special Issue Hepatitis B Virus: Its Life Cycle and the Therapeutic Targets)

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Abstract

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 × 10¹¹ enveloped, DNA-containing HBV particles/mL each week), and is with minimal equipment easily adaptable to most laboratory settings.

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

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MDPI and ACS Style

Wettengel, J.M.; Linden, B.; Esser, K.; Laue, M.; Burwitz, B.J.; Protzer, U. Rapid and Robust Continuous Purification of High-Titer Hepatitis B Virus for In Vitro and In Vivo Applications. Viruses 2021, 13, 1503. https://doi.org/10.3390/v13081503

AMA Style

Wettengel JM, Linden B, Esser K, Laue M, Burwitz BJ, Protzer U. Rapid and Robust Continuous Purification of High-Titer Hepatitis B Virus for In Vitro and In Vivo Applications. Viruses. 2021; 13(8):1503. https://doi.org/10.3390/v13081503

Chicago/Turabian Style

Wettengel, Jochen M., Bianca Linden, Knud Esser, Michael Laue, Benjamin J. Burwitz, and Ulrike Protzer. 2021. "Rapid and Robust Continuous Purification of High-Titer Hepatitis B Virus for In Vitro and In Vivo Applications" Viruses 13, no. 8: 1503. https://doi.org/10.3390/v13081503

APA Style

Wettengel, J. M., Linden, B., Esser, K., Laue, M., Burwitz, B. J., & Protzer, U. (2021). Rapid and Robust Continuous Purification of High-Titer Hepatitis B Virus for In Vitro and In Vivo Applications. Viruses, 13(8), 1503. https://doi.org/10.3390/v13081503

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

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