Sars-cov-2 production in a scalable high cell density bioreactor
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- SARS-CoV-2 Production in a Scalable High Cell Density Bioreactor
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The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has demonstrated the value of pursuing different vaccine strategies. Vaccines based on whole viruses, a widely used vaccine technology, depend on efficient virus production. This study aimed to establish SARSCoV-2 production in the scalable packed-bed CelCradle™ 500-AP bioreactor. CelCradle™ 500-AP bottles with 0.5 L working volume and 5.5 g BioNOC™ II carriers were seeded with 1.5 × 108 Vero (WHO) cells, approved for vaccine production, in animal component-free medium and infected at a multiplicity of infection of 0.006 at a total cell number of 2.2–2.5 × 109 cells/bottle seven days post cell seeding. Among several tested conditions, two harvests per day and a virus production temperature of 33◦C resulted in the highest virus yield with a peak SARS-CoV-2 infectivity titer of 7.3 log10 50% tissue culture infectious dose (TCID50)/mL at 72 h post-infection. Six harvests had titers of ≥6.5 log10 TCID50/mL, and a total of 10.5 log10 TCID50 were produced in ~5 L. While trypsin was reported to enhance virus spread in cell culture, addition of 0.5% recombinant trypsin after infection did not improve virus yields. Overall, we demonstrated successful animal component-free production of SARS-CoV-2 in well-characterized Vero (WHO) cells in a scalable packed-bed bioreactor.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 706 |
| Tidsskrift | Vaccines |
| Vol/bind | 9 |
| Udgave nummer | 7 |
| ISSN | 2076-393X |
| DOI | |
| Status | Udgivet - 2021 |
Bibliografisk note
Funding Information:
This study was supported by grants from the Candys Foundation (A.O., A.F.P., L.V.P., J.B., J.M.G.), the Danish Agency for Science and Higher Education (S.R., J.B.), the Danish Cancer Society (J.B.), Hvidovre Hospital Research Foundation (C.R.D.H., J.M.G.), Independent Research Fund Denmark (DFF) Medical Sciences (J.B., J.M.G.), Innovation Fund Denmark (J.B., J.M.G.), the L?ge Sofus Carl Emil Friis og Hustru Olga Doris Friis? Foundation (J.M.G.), Mauritzen La Fontaine Fonden (J.B., J.M.G.), the Novo Nordisk Foundation (J.B.), the Region H Foundation (J.B., J.M.G.), the Toyota Foundation (A.O., J.M.G.), the Weimann Foundation (U.F.). Acknowledgments: We thank Lotte Mikkelsen, Anna-Louise S?rensen, and Pia Pedersen (Copenhagen University Hospital, Hvidovre) for laboratory assistance and Bjarne ?. Lindhardt (Copenhagen University Hospital, Hvidovre) and Carsten Geisler (University of Copenhagen) for their support. We thank Nuvonis Technologies GmbH (Vienna, Austria) for providing Vero (WHO) cells (ECACC no 88020401) and corresponding protocols, and M. Wolschek of Nuvonis for helpful discussion. We thank Jean Dubuisson (J.D.), University of Lille, for providing Vero E6 cells.
Funding Information:
Funding: This study was supported by grants from the Candys Foundation (A.O., A.F.P., L.V.P., J.B., J.M.G.), the Danish Agency for Science and Higher Education (S.R., J.B.), the Danish Cancer Society (J.B.), Hvidovre Hospital Research Foundation (C.R.D.H., J.M.G.), Independent Research Fund Denmark (DFF) Medical Sciences (J.B., J.M.G.), Innovation Fund Denmark (J.B., J.M.G.), the Læge Sofus Carl Emil Friis og Hustru Olga Doris Friis’ Foundation (J.M.G.), Mauritzen La Fontaine Fonden (J.B., J.M.G.), the Novo Nordisk Foundation (J.B.), the Region H Foundation (J.B., J.M.G.), the Toyota Foundation (A.O., J.M.G.), the Weimann Foundation (U.F.).
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
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