TY - JOUR
T1 - Sars-cov-2 production in a scalable high cell density bioreactor
AU - Offersgaard, Anna
AU - Hernandez, Carlos Rene Duarte
AU - Pihl, Anne Finne
AU - Costa, Rui
AU - Venkatesan, Nandini Prabhakar
AU - Lin, Xiangliang
AU - Van Pham, Long
AU - Feng, Shan
AU - Fahnøe, Ulrik
AU - Scheel, Troels Kasper Høyer
AU - Ramirez, Santseharay
AU - Reichl, Udo
AU - Bukh, Jens
AU - Genzel, Yvonne
AU - Gottwein, Judith Margarete
N1 - Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021
Y1 - 2021
N2 - 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.
AB - 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.
KW - Animal component-free
KW - CelCradle
KW - COVID-19
KW - Inactivated vaccine
KW - Packed-bed
KW - Scalable bioreactor
KW - Severe acute respiratory syndrome coronavirus 2
KW - Vero cells
KW - Whole virus vaccine
U2 - 10.3390/vaccines9070706
DO - 10.3390/vaccines9070706
M3 - Journal article
C2 - 34209694
AN - SCOPUS:85109955506
VL - 9
JO - Vaccines
JF - Vaccines
SN - 2076-393X
IS - 7
M1 - 706
ER -