TY - JOUR
T1 - High‐Titer Hepatitis C Virus Production in a Scalable Single‐Use High Cell Density Bioreactor
AU - Offersgaard, Anna
AU - Duarte Hernandez, Carlos Rene
AU - Finne Pihl, Anne
AU - Prabhakar Venkatesan, Nandini
AU - Krarup, Henrik
AU - Lin, Xiangliang
AU - Reichl, Udo
AU - Bukh, Jens
AU - Genzel, Yvonne
AU - Gottwein, Judith Margarete
N1 - Publisher Copyright:
© 2022 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2022/2
Y1 - 2022/2
N2 - Hepatitis C virus (HCV) infections pose a major public health burden due to high chronicity rates and associated morbidity and mortality. A vaccine protecting against chronic infection is not available but would be important for global control of HCV infections. In this study, cell culture‐based HCV production was established in a packed‐bed bioreactor (CelCradle™) aiming to further the development of an inactivated whole virus vaccine and to facilitate virological and immunological studies requiring large quantities of virus particles. HCV was produced in human hepatoma‐derived Huh7.5 cells maintained in serum‐free medium on days of virus harvesting. Highest virus yields were obtained when the culture was maintained with two medium exchanges per day. However, increasing the total number of cells in the culture vessel negatively impacted infectivity titers. Peak infectivity titers of up to 7.2 log10 focus forming units (FFU)/mL, accumulated virus yields of up to 5.9 × 1010 FFU, and a cell specific virus yield of up to 41 FFU/cell were obtained from one CelCradle™. CelCradle™‐derived and T flask‐derived virus had similar characteristics regarding neutralization sensitivity and buoyant density. This packed‐bed tide‐motion system is available with larger vessels and may thus be a promising platform for large‐scale HCV production.
AB - Hepatitis C virus (HCV) infections pose a major public health burden due to high chronicity rates and associated morbidity and mortality. A vaccine protecting against chronic infection is not available but would be important for global control of HCV infections. In this study, cell culture‐based HCV production was established in a packed‐bed bioreactor (CelCradle™) aiming to further the development of an inactivated whole virus vaccine and to facilitate virological and immunological studies requiring large quantities of virus particles. HCV was produced in human hepatoma‐derived Huh7.5 cells maintained in serum‐free medium on days of virus harvesting. Highest virus yields were obtained when the culture was maintained with two medium exchanges per day. However, increasing the total number of cells in the culture vessel negatively impacted infectivity titers. Peak infectivity titers of up to 7.2 log10 focus forming units (FFU)/mL, accumulated virus yields of up to 5.9 × 1010 FFU, and a cell specific virus yield of up to 41 FFU/cell were obtained from one CelCradle™. CelCradle™‐derived and T flask‐derived virus had similar characteristics regarding neutralization sensitivity and buoyant density. This packed‐bed tide‐motion system is available with larger vessels and may thus be a promising platform for large‐scale HCV production.
KW - CelCradle™
KW - HCV vaccine development
KW - High cell density cell culture
KW - High‐titer HCV production
KW - Huh7.5 cells
KW - Inactivated virus vaccine
KW - Packed‐bed bioreactor
KW - Whole virus vaccine
UR - http://www.scopus.com/inward/record.url?scp=85124391880&partnerID=8YFLogxK
U2 - 10.3390/vaccines10020249
DO - 10.3390/vaccines10020249
M3 - Journal article
AN - SCOPUS:85124391880
SN - 2076-393X
VL - 10
JO - Vaccines
JF - Vaccines
IS - 2
M1 - 249
ER -