@article{b18fd78e9106450a85dc996f04dbaa1f,
title = "Production of rVSV-ZEBOV in serum-free suspension culture of HEK 293SF cells",
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",
author = "G{\'e}linas, {Jean Fran{\c c}ois} and Hiva Azizi and Sascha Kiesslich and St{\'e}phane Lanthier and Jannie Perdersen and Chahal, {Parminder S.} and Sven Ansorge and Gary Kobinger and R{\'e}nald Gilbert and Kamen, {Amine A.}",
note = "Funding Information: We thank M{\'e}lanie Leclerc and Nathalie Coulombe from the National Research Council Canada who performed the virus purification for the samples used in animal studies and Julien Robitaille, also from the National Research Council Canada , who performed data extraction for the bioreactor. This research was funded by the Canadian Institutes of Health Research Grant OVV 152411 . JFG is funded by a Natural Sciences and Engineering Research Council of Canada (NSERC) postdoctoral fellowship. SK is funded by a doctoral scholarship from the Fonds de Recherche du Qu{\'e}bec – Sant{\'e} (FRQS). The funding sources had no involvement in the study design, in the collection, analysis and interpretation of data, in the writing of the report or in the decision to submit the article for publication. Funding Information: We thank M{\'e}lanie Leclerc and Nathalie Coulombe from the National Research Council Canada who performed the virus purification for the samples used in animal studies and Julien Robitaille, also from the National Research Council Canada, who performed data extraction for the bioreactor. This research was funded by the Canadian Institutes of Health Research Grant OVV 152411. JFG is funded by a Natural Sciences and Engineering Research Council of Canada (NSERC) postdoctoral fellowship. SK is funded by a doctoral scholarship from the Fonds de Recherche du Qu{\'e}bec – Sant{\'e} (FRQS). The funding sources had no involvement in the study design, in the collection, analysis and interpretation of data, in the writing of the report or in the decision to submit the article for publication. JP performed the plasmid cloning and the viral rescue described in the supplementary methods. JFG, RG and AAK designed the cell culture experiments and JFG performed them. JFG and SK designed the dPCR protocol and SK performed this assay. JFG and SL performed the bioreactor production. PSC designed and supervised the purification process. HA designed and performed the animal studies. SA, GK, RG and AAK provided material and financial support. JFG wrote the manuscript and all authors revised the final version. Publisher Copyright: {\textcopyright} 2019",
year = "2019",
month = oct,
day = "16",
doi = "10.1016/j.vaccine.2019.09.044",
language = "English (US)",
volume = "37",
pages = "6624--6632",
journal = "Vaccine",
issn = "0264-410X",
publisher = "Elsevier BV",
number = "44",
}