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SARS-CoV-2 Production in a Scalable High Cell Density Bioreactor

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@article{7e3e956b6d8c40f59820ee7884b6181d,
title = "SARS-CoV-2 Production in a Scalable High Cell Density Bioreactor",
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 SARS-CoV-2 production in the scalable packed-bed CelCradleTM 500-AP bioreactor. CelCradleTM 500-AP bottles with 0.5 L working volume and 5.5 g BioNOC{\texttrademark} 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.",
keywords = "Animal component-free, CelCradle, COVID-19, Inactivated vaccine, Packed-bed, Scalable bioreactor, Severe acute respiratory syndrome coronavirus 2, Vero cells, Whole virus vaccine",
author = "Anna Offersgaard and {Duarte Hernandez}, {Carlos Rene} and Pihl, {Anne Finne} and Rui Costa and Venkatesan, {Nandini Prabhakar} and Xiangliang Lin and {Van Pham}, Long and Shan Feng and Ulrik Fahn{\o}e and Scheel, {Troels Kasper H{\o}yer} and Santseharay Ramirez and Udo Reichl and Jens Bukh and Yvonne Genzel and Gottwein, {Judith Margarete}",
year = "2021",
month = jun,
day = "29",
doi = "10.3390/vaccines9070706",
language = "English",
volume = "9",
journal = "Vaccines",
issn = "1554-8600",
publisher = "Landes Bioscience",
number = "7",

}

RIS

TY - JOUR

T1 - SARS-CoV-2 Production in a Scalable High Cell Density Bioreactor

AU - Offersgaard, Anna

AU - Duarte Hernandez, Carlos Rene

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

PY - 2021/6/29

Y1 - 2021/6/29

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 SARS-CoV-2 production in the scalable packed-bed CelCradleTM 500-AP bioreactor. CelCradleTM 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 SARS-CoV-2 production in the scalable packed-bed CelCradleTM 500-AP bioreactor. CelCradleTM 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

UR - http://www.scopus.com/inward/record.url?scp=85109955506&partnerID=8YFLogxK

U2 - 10.3390/vaccines9070706

DO - 10.3390/vaccines9070706

M3 - Journal article

C2 - 34209694

VL - 9

JO - Vaccines

JF - Vaccines

SN - 1554-8600

IS - 7

M1 - 706

ER -

ID: 66572975