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Production of yellow fever VLPs by perfusion cultivation of stable recombinant HEK293 cells
  • +2
  • Renata Alvim,
  • Túlio Lima,
  • Jerson Silva,
  • Guilherme Oliveira,
  • Leda Castilho
Renata Alvim
Federal University of Rio de Janeiro
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Túlio Lima
Federal University of Rio de Janeiro
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Jerson Silva
Federal University of Rio de Janeiro
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Guilherme Oliveira
Federal University of Rio de Janeiro
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Leda Castilho
Federal University of Rio de Janeiro
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Abstract

Yellow fever (YF) is a life-threatening viral disease endemic in large areas of Africa and Latin America. Although there is a very efficacious vaccine since the 1930s, YF still causes 29,000-60,000 annual deaths. During recent YF outbreaks there were issues of vaccine shortage due to limited supply of the current egg-derived vaccine; rare but fatal vaccine adverse effects occurred; and cases were imported to Asia, where the mosquito vector circulates and where local transmission could potentially start. In this work, we investigated the production of YF virus-like particles (VLPs) using suspension-adapted stably-transfected HEK293 cells. In order to develop an intensified process, we combined two strategies: the use of sequential FACS rounds to enrich the stable cell pool in terms of high producers, and the use of perfusion processes. At first, shaken tube experiments revealed that FACS enrichment of the cell pool allowed doubling VLP production, and that in pseudoperfusion cultures (with daily medium exchange) lasting 14 days VLP production increased by 8.3 fold as compared to batch cultures lasting 11 days. When true perfusion cultures were carried out in bioreactors, the use of an inclined cell settler as cell retention device showed operational advantages as compared to an ATF system.

Peer review status:ACCEPTED

05 Oct 2020Submitted to Biotechnology and Bioengineering
07 Oct 2020Submission Checks Completed
07 Oct 2020Assigned to Editor
16 Nov 2020Reviewer(s) Assigned
04 Jan 2021Review(s) Completed, Editorial Evaluation Pending
04 Jan 2021Editorial Decision: Revise Minor
08 Apr 20211st Revision Received
09 Apr 2021Submission Checks Completed
09 Apr 2021Assigned to Editor
14 Jun 2021Review(s) Completed, Editorial Evaluation Pending
14 Jun 2021Editorial Decision: Accept