Optimization of Early Steps in Oncolytic Adenovirus ONCOS-401 Production in T-175 and HYPERFlasks

International Journal of Molecular Sciences

Mariangela Garofalo¹  Vincenzo Cerullo²  Magnus Jaderberg²  Anne-SophieW Møller³  Antti Vuolanto⁴  Lukasz Kuryk⁵  Sari Pesonen⁶ 

[1] D, 00180 Helsinki, Finland;Drug Research Program, ImmunoVirothearpy Lab, Faculty of Pharmacy, University of Helsinki, 00014 Helsinki, Finland;Targovax ASA, Clinical Science, 0283 Oslo, Norway;Targovax Oy, CMC, 00180 Helsinki, Finland;Targovax Oy, Clinical Science, 00180 Helsinki, Finland;;Targovax Oy, R&

关键词: oncolytic adenovirus;    CD40L;    productivity;    benzonase;    manufacturing;    optimization;    cancer;    MOI;    harvesting time;    virus productivity;   

DOI  :  10.3390/ijms20030621

来源: DOAJ

【 摘 要 】

Oncolytic adenoviruses can trigger lysis of tumor cells, induce an antitumor immune response, bypass classical chemotherapeutic resistance strategies of tumors, and provide opportunities for combination strategies. A major challenge is the development of scalable production methods for viral seed stocks and sufficient quantities of clinical grade viruses. Because of promising clinical signals in a compassionate use program (Advanced Therapy Access Program) which supported further development, we chose the oncolytic adenovirus ONCOS-401 as a testbed for a new approach to scale up. We found that the best viral production conditions in both T-175 flasks and HYPERFlasks included A549 cells grown to 220,000 cells/cm² (80% confluency), with ONCOS-401 infection at 30 multiplicity of infection (MOI), and an incubation period of 66 h. The Lysis A harvesting method with benzonase provided the highest viral yield from both T-175 and HYPERFlasks (10,887 ± 100 and 14,559 ± 802 infectious viral particles/cell, respectively). T-175 flasks and HYPERFlasks produced up to 2.1 × 10⁹ ± 0.2 and 1.75 × 10⁹ ± 0.08 infectious particles of ONCOS-401 per cm² of surface area, respectively. Our findings suggest a suitable stepwise process that can be applied to optimizing the initial production of other oncolytic viruses.

【 授权许可】

Unknown