Production of rVSV-ZEBOV in serum-free suspension culture of HEK 293SF cells
| Autor: | Jannie Perdersen, Parminder S. Chahal, Stéphane Lanthier, Gary P. Kobinger, Sascha Kiesslich, Hiva Azizi, Jean-François Gélinas, Amine Kamen, Sven Ansorge, Rénald Gilbert |
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| Rok vydání: | 2019 |
| Předmět: |
rVSV-ZEBOV
030231 tropical medicine serum-free HEK 293SF Antibodies Viral medicine.disease_cause Suspension culture Virus Mice 03 medical and health sciences Bioreactors 0302 clinical medicine Immune system Serum free vaccine Vaccines DNA medicine Bioreactor suspension Animals Humans 030212 general & internal medicine Ebola Vaccines Ebola virus General Veterinary General Immunology and Microbiology Chemistry HEK 293 cells Public Health Environmental and Occupational Health Vesiculovirus Hemorrhagic Fever Ebola Ebolavirus Virology 3. Good health Disease Models Animal Titer HEK293 Cells Infectious Diseases Batch Cell Culture Techniques Ebola Molecular Medicine Female Immunization production |
| Zdroj: | Vaccine. 37:6624-6632 |
| ISSN: | 0264-410X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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