Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5537358 | Vaccine | 2016 | 8 Pages |
Abstract
The selection of a cell substrate is a critical step for the development and manufacturing of a viral vaccine candidate. Several parameters such as cell susceptibility and permissiveness to the viral pathogens but also performance in terms of viral antigens quality and production yields are important considerations when identifying the ideal match between a viral vaccine and cell substrate. The modified vaccinia virus Ankara (MVA) is a replication-deficient viral vector that holds great promise as a vaccine platform, however only limited cell substrates have been tested or are available for industrialization. Here we evaluate the duck embryo-derived EB66® cell line as potential cell substrate for MVA production. To this end, we used two recombinant MVA constructs and demonstrated that EB66® cells are propagating the tested MVA viruses very efficiently, while preserving viral attenuation and transgene expression for up to 20 serial passages. Furthermore we developed upstream and downstream processes that enable industrialization of the virus production. In conclusion, we showed that EB66® cells can be used as potent cell substrate for MVA-based vaccines and represent therefore an attractive alternative for vaccine production.
Keywords
GMPHost cell DNATCID50CEVMGBEMEMCEFMRCHCDHCPHDFqPCRFBSGFPDMEMDNAMOITffdeoxyribonucleic acidModified vaccinia virus AnkaraEnzyme-linked immunosorbent assayELISATangential flow filtrationGood Manufacturing PracticeVaccine productionEagle’s Minimum Essential Mediumfoetal bovine serumMedical Research CouncilManufacturing processChicken embryo fibroblasthuman dermal fibroblastMVADulbecco’s modified eagle’s mediumHepatitis C virusHCVquantitative polymerase chain reactionhost cell proteingreen fluorescent proteinPES, Polyethersulfonemultiplicity of infectionMinor groove binder
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Authors
Arnaud Léon, Anne-Laure David, Brice Madeline, Laurence Guianvarc'h, Elodie Dureau, Patrick Champion-Arnaud, Matthias Hebben, Thierry Huss, Benoît Chatrenet, Klaus Schwamborn,