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

This study demonstrates the feasibility of large-scale production of murine polyomavirus VP1 protein in recombinant Escherichia colt 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 600 nm of 20 60 or 100) GST-VP1 production was highest when the culture was Induced at a cell density of 00 60 with volumetric yield reaching 438 g L-1 in 31 h 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 colt 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 (C) 2010 Elsevier B V All rights reserved