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Vaccine. 2019 Oct 16;37(44):6624-6632. doi: 10.1016/j.vaccine.2019.09.044. Epub 2019 Sep 20.

Production of rVSV-ZEBOV in serum-free suspension culture of HEK 293SF cells.

Author information

1
Department of Bioengineering, McGill University, Montreal, QC, Canada.
2
Centre de Recherche en Infectiologie, Centre Hospitalier Universitaire de Québec, Université Laval, Quebec City, QC, Canada.
3
Human Health Therapeutics, National Research Council Canada, Montreal, QC, Canada.
4
Department of Bioengineering, McGill University, Montreal, QC, Canada; Human Health Therapeutics, National Research Council Canada, Montreal, QC, Canada.
5
Department of Bioengineering, McGill University, Montreal, QC, Canada. Electronic address: amine.kamen@mcgill.ca.

Abstract

Ebola virus disease is an urgent international priority. Promising results for several vaccine candidates have been reported in non-human primate studies and clinical trials with the most promising being the rVSV-ZEBOV vaccine. In this study, we sought to produce rVSV-ZEBOV in HEK 293SF cells in suspension and serum-free media. The purpose of this study was to establish a process using the HEK 293SF production platform, optimise the production titre, demonstrate scalability and the efficiency of the generated material to elicit an immune reaction in an animal model. Critical process parameters were evaluated to maximize production yield and process robustness and the following operating conditions: 1-2 × 106 cells/mL grown in HyClone HyCell TransFx-H media infected at an MOI of 0.001 with a temperature shift to 34 °C during the production phase and a harvest of the product after 48 h. Using these conditions, scalability in a 3.5 L controlled bioreactor was shown reaching a titre of 1.19 × 108 TCID50/mL at the peak of production, the equivalent of 4165 doses of vaccine per litre. The produced virus was shown to be thermostable in the culture media and, when concentrated, purified and administered to mice, demonstrated the ability to induce a ZEBOV-specific immune response.

KEYWORDS:

Ebola; HEK 293SF; Production; Serum-free; Suspension; Vaccine; rVSV-ZEBOV

PMID:
31548015
DOI:
10.1016/j.vaccine.2019.09.044
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