Sherry, L orcid.org/0000-0002-4367-772X, Grehan, K orcid.org/0000-0003-3976-756X, Snowden, JS orcid.org/0000-0001-7857-0634 et al. (4 more authors) (2020) Comparative Molecular Biology Approaches for the Production of Poliovirus Virus-Like Particles Using Pichia pastoris. mSphere, 5 (2). e00838-19. ISSN 2379-5042
Abstract
For enteroviruses such as poliovirus (PV), empty capsids, which are antigenically indistinguishable from mature virions, are produced naturally during viral infection. The production of such capsids recombinantly, in heterologous systems such as yeast, have great potential as virus-like particle (VLP) vaccine candidates. Here, using PV as an exemplar, we show the production of VLPs in Pichia pastoris by coexpression of the structural precursor protein P1 and the viral protease 3CD. The level of expression of the potentially cytotoxic protease relative to that of the P1 precursor was modulated by three different approaches: expression of the P1 precursor and protease from different transcription units, separation of the P1 and protease proteins using the Thosea asigna virus (TaV) 2A translation interruption sequence, or separation of the P1 and protease-coding sequences by an internal ribosome entry site sequence from Rhopalosiphum padi virus (RhPV). We also investigate the antigenicity of VLPs containing previously characterized mutations when produced in Pichia. Finally, using transmission electron microscopy and two-dimensional classification, we show that Pichia-derived VLPs exhibited the classical icosahedral capsid structure displayed by enteroviruses.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2020 Sherry et al. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International license. |
Keywords: | enterovirus, poliovirus, virus-like particle, Pichia pastoris |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Molecular Virology (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 12 Mar 2020 11:42 |
Last Modified: | 25 Jun 2023 22:11 |
Status: | Published |
Publisher: | American Society for Microbiology |
Identification Number: | 10.1128/msphere.00838-19 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:158344 |