title: Microbial production of virus-like particle vaccine protein at gram-per-litre levels.
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 600 nm 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 4.38 gL⁻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.
authors.affiliation: The University of Queensland, Centre for Biomolecular Engineering, Australian Institute for Bioengineering and Nanotechnology, St Lucia, QLD 4072, Australia.
authors.forename: Mervyn W O
authors.initials: MW
authors.lastname: Liew
authors.forename: Aravindan
authors.initials: A
authors.lastname: Rajendran
authors.forename: Anton P J
authors.initials: AP
authors.lastname: Middelberg
mesh_terms: D018533:Biomass; D019149:Bioreactors; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002452:Cell Count; D049109:Cell Proliferation; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D031521:Fractionation, Field Flow; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D046529:Microscopy, Electron, Transmission; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D058425:Vaccines, Virus-Like Particle / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C442540:VP1 protein, Poliovirus; D058425:Vaccines, Virus-Like Particle; D005982:Glutathione Transferase