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

Liew, Mervyn W. O., Rajendran, Aravindan and Middelberg, Anton P. J. (2010) Microbial production of virus-like particle vaccine protein at gram-per-litre levels. Journal of Biotechnology, 150 2: 224-231. doi:10.1016/j.jbiotec.2010.08.010


Author Liew, Mervyn W. O.
Rajendran, Aravindan
Middelberg, Anton P. J.
Title Microbial production of virus-like particle vaccine protein at gram-per-litre levels
Journal name Journal of Biotechnology   Check publisher's open access policy
ISSN 0168-1656
1873-4863
Publication date 2010-10
Sub-type Article (original research)
DOI 10.1016/j.jbiotec.2010.08.010
Volume 150
Issue 2
Start page 224
End page 231
Total pages 8
Place of publication Amsterdam, Netherlands
Publisher Elsevier BV
Collection year 2011
Language eng
Formatted 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 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. 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.
Copyright © 2010 Elsevier B.V. All rights reserved.
Keyword Virus like particles
VP1
Heterologous gene expression
Fed batch cultivation
pH-stat
Escherichia coli
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 28 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 30 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Sun, 12 Dec 2010, 00:06:05 EST