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

Research output: Contribution to journalJournal articleResearchpeer-review

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

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)406-12
Number of pages7
Publication statusPublished - Feb 2013

Bibliographical note

Copyright © 2012 Elsevier B.V. All rights reserved.

    Research areas

  • Animals, Biotechnology/methods, Capsid/immunology, Cell Line, Cysteine Endopeptidases/biosynthesis, Down-Regulation, Foot-and-Mouth Disease Virus/genetics, Gene Expression, Insecta, Technology, Pharmaceutical/methods, Viral Proteins/biosynthesis, Viral Vaccines/genetics

ID: 257916853