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SARS-CoV-2 Production in a Scalable High Cell Density Bioreactor #MMPMID34209694
Offersgaard A; Duarte Hernandez CR; Pihl AF; Costa R; Venkatesan NP; Lin X; Van Pham L; Feng S; Fahnoe U; Scheel TKH; Ramirez S; Reichl U; Bukh J; Genzel Y; Gottwein JM
Vaccines (Basel) 2021[Jun]; 9 (7): ? PMID34209694show ga
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 SARS-CoV-2 production in the scalable packed-bed CelCradle(TM) 500-AP bioreactor. CelCradle(TM) 500-AP bottles with 0.5 L working volume and 5.5 g BioNOC II carriers were seeded with 1.5 x 10(8) 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 x 10(9) cells/bottle seven days post cell seeding. Among several tested conditions, two harvests per day and a virus production temperature of 33 degrees C resulted in the highest virus yield with a peak SARS-CoV-2 infectivity titer of 7.3 log(10) 50% tissue culture infectious dose (TCID(50))/mL at 72 h post-infection. Six harvests had titers of >/=6.5 log(10) TCID(50)/mL, and a total of 10.5 log(10) TCID(50) 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.
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Vaccines+(Basel) 2021 ; 9 (7): ?
