Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity

Claudine Porta, Xiaodong Xu, Silvia Loureiro, Saravanan Paramasivam, Junyuan Ren, Tara Al-Khalil, Alison Burman, Terry Jackson, Graham Belsham, Stephen Curry, George P. Lomonossoff, Satya Parida, David Paton, Yanmin Li, Ginette Wilsden, Nigel Ferris, Ray Owens, Abhay Kotecha, Elizabeth Fry, David I. StuartBryan Charleston, Ian M. Jones

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    Foot-and-mouth disease virus (FMDV) is a significant economically and distributed globally pathogen of Artiodactyla. Current vaccines are chemically inactivated whole virus particles that require large-scale virus growth in strict bio-containment with the associated risks of accidental release or incomplete inactivation. Non-infectious empty capsids are structural mimics of authentic particles with no associated risk and constitute an alternate vaccine candidate. Capsids self-assemble from the processed virus structural proteins, VP0, VP3 and VP1, which are released from the structural protein precursor P1-2A by the action of the virus-encoded 3C protease. To date recombinant empty capsid assembly has been limited by poor expression levels, restricting the development of empty capsids as a viable vaccine. Here expression of the FMDV structural protein precursor P1-2A in insect cells is shown to be efficient but linkage of the cognate 3C protease to the C-terminus reduces expression significantly. Inactivation of the 3C enzyme in a P1-2A-3C cassette allows expression and intermediate levels of 3C activity resulted in efficient processing of the P1-2A precursor into the structural proteins which assembled into empty capsids. Expression was independent of the insect host cell background and leads to capsids that are recognised as authentic by a range of anti-FMDV bovine sera suggesting their feasibility as an alternate vaccine.
    Original languageEnglish
    JournalJournal of Virological Methods
    Issue number2
    Pages (from-to)406-412
    Publication statusPublished - 2013


    • Foot-and-mouth disease virus
    • Recombinant baculovirus
    • Empty capsids
    • Protein processing
    • Frameshift
    • 3C protease
    • Vaccine


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