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Open AccessArticle

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

1
Targovax Oy, Clinical Science, 00180 Helsinki, Finland
2
National Institute of Public Health–National Institute of Hygiene, Department of Virology, 00-0791 Warsaw, Poland
3
Drug Research Program, ImmunoVirothearpy Lab, Faculty of Pharmacy, University of Helsinki, 00014 Helsinki, Finland
4
Targovax ASA, Clinical Science, 0283 Oslo, Norway
5
Targovax Oy, CMC, 00180 Helsinki, Finland
6
Targovax Oy, R&D, 00180 Helsinki, Finland
7
Department of Oncology and Hemato-Oncology, Center of Excellence on Neurodegenerative Diseases, University of Milan, 20133 Milan, Italy
*
Author to whom correspondence should be addressed.
Current address: Herantis Pharma Plc, 02600 Espoo, Finland.
Current address: Valo Therapeutics Ltd., 00100 Helsinki, Finland.
Int. J. Mol. Sci. 2019, 20(3), 621; https://doi.org/10.3390/ijms20030621
Received: 20 December 2018 / Revised: 15 January 2019 / Accepted: 29 January 2019 / Published: 31 January 2019
(This article belongs to the Special Issue Tumor Targeting Therapy and Selective Killing 2018)
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. View Full-Text
Keywords: oncolytic adenovirus; CD40L; productivity; benzonase; manufacturing; optimization; cancer; MOI; harvesting time; virus productivity oncolytic adenovirus; CD40L; productivity; benzonase; manufacturing; optimization; cancer; MOI; harvesting time; virus productivity
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Kuryk, L.; Møller, A.-S.W.; Vuolanto, A.; Pesonen, S.; Garofalo, M.; Cerullo, V.; Jaderberg, M. Optimization of Early Steps in Oncolytic Adenovirus ONCOS-401 Production in T-175 and HYPERFlasks. Int. J. Mol. Sci. 2019, 20, 621.

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