High-Titer Hepatitis C Virus Production in a Scalable Single-Use High Cell Density Bioreactor

• Published in: Vaccines (Basel) Vol. 10; no. 2; p. 249
• Main Authors: Offersgaard, Anna, Duarte Hernandez, Carlos Rene, Pihl, Anne Finne, Venkatesan, Nandini Prabhakar, Krarup, Henrik, Lin, Xiangliang, Reichl, Udo, Bukh, Jens, Genzel, Yvonne, Gottwein, Judith Margarete
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 07-02-2022; MDPI
• Subjects: Antibodies; Bioreactors; CelCradle; Cell culture; Cell density; Chronic infection; Density; Efficiency; Experiments; Good Manufacturing Practice; Hepatitis; Hepatitis C; Hepatoma; high cell density cell culture; Huh7.5 cells; Immunization; Immunology; inactivated virus vaccine; Infections; Infectivity; Morbidity; Motion systems; Neutralization; packed-bed bioreactor; Proteins; Public health; Serum-free medium; Severe acute respiratory syndrome coronavirus 2; Vaccines; Vessels; Viruses; whole virus vaccine; Singapore; Huh7.5 cells; high cell density cell culture; packed-bed bioreactor; inactivated virus vaccine; CelCradle; high-titer HCV production; HCV vaccine development; whole virus vaccine
• ISSN: 2076-393X; 2076-393X
• DOI: 10.3390/vaccines10020249

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 log
focus forming units (FFU)/mL, accumulated virus yields of up to 5.9 × 10
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.