Microbial production of virus-like particle vaccine protein at gram-per-litre levels

Mervyn W O Liew, Aravindan Rajendran, Anton P J Middelberg

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44 Scopus citations

Abstract

This study demonstrates the feasibility of large-scale production of murine polyomavirus VP1 protein in recombinant Escherichia coli as pentamers which are able to subsequently self-assemble in vitro into virus-like particles (VLPs). High-cell-density pH-stat fed-batch cultivation was employed to produce glutathione-S-transferase (GST)-VP1 fusion protein in soluble form. The expression of recombinant VP1 was induced with IPTG at different cell optical densities (OD at 600nm of 20, 60 or 100). GST-VP1 production was highest when the culture was induced at a cell density of OD 60, with volumetric yield reaching 4.38gL-1 in 31h, which we believe is the highest volumetric productivity for viral capsid protein reported to date. The induction cell density is shown to have a significant effect on the overall volumetric yield of recombinant VP1 and on final cell density, but not on VLP quality. VP1 yield was enhanced 15-fold by scaling-up from shake flask to pH-stat fed-batch cultivation in a bioreactor. Although numerous studies have expressed structural viral protein in E. coli, we believe this is the first report of translation to bioreactors yielding gram-per-litre levels. This VLP production technology overcomes major drawbacks associated with eukaryotic cell-based vaccine production technologies, and propounds the scope for large-scale commercially viable E. coli based VLP production by significantly reducing vaccine production time and cost.

Original languageEnglish (US)
Pages (from-to)224-231
Number of pages8
JournalJournal of Biotechnology
Volume150
Issue number2
DOIs
StatePublished - Oct 2010

Bibliographical note

Funding Information:
Mervyn W.O. Liew acknowledges funding from the Ministry of Higher Education – Universiti Sains Malaysia in the form of a post-graduate fellowship. Aravindan Rajendran acknowledges support from the Australian Government through an Endeavour Research Fellowship. Anton Middelberg acknowledges support from the Queensland Government in the form of a Smart Futures Premier's Fellowship.

Copyright:
Copyright 2011 Elsevier B.V., All rights reserved.

Keywords

  • Escherichia coli
  • Fed-batch cultivation
  • Heterologous gene expression
  • PH-stat
  • VP1
  • Virus-like particles

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