Sars-cov-2 production in a scalable high cell density bioreactor

Anna Offersgaard, Carlos Rene Duarte Hernandez, Anne Finne Pihl, Rui Costa, Nandini Prabhakar Venkatesan, Xiangliang Lin, Long Van Pham, Shan Feng, Ulrik Fahnøe, Troels Kasper Høyer Scheel, Santseharay Ramirez, Udo Reichl, Jens Bukh, Yvonne Genzel, Judith Margarete Gottwein*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

19 Citations (Scopus)
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Abstract

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 33C 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.

Original languageEnglish
Article number706
JournalVaccines
Volume9
Issue number7
ISSN2076-393X
DOIs
Publication statusPublished - 2021

Bibliographical note

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

  • Animal component-free
  • CelCradle
  • COVID-19
  • Inactivated vaccine
  • Packed-bed
  • Scalable bioreactor
  • Severe acute respiratory syndrome coronavirus 2
  • Vero cells
  • Whole virus vaccine

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