Next Article in Journal
Novel Zebrafish Mono-α2,8-sialyltransferase (ST8Sia VIII): An Evolutionary Perspective of α2,8-Sialylation
Previous Article in Journal
Transcriptome Analysis of Improved Wool Production in Skin-Specific Transgenic Sheep Overexpressing Ovine β-Catenin
Previous Article in Special Issue
Specific Antibody Fragment Ligand Traps Blocking FGF1 Activity
Article Menu
Issue 3 (February-1) cover image

Export Article

Open AccessArticle
Int. J. Mol. Sci. 2019, 20(3), 621; https://doi.org/10.3390/ijms20030621

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.
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)
  |  
PDF [1374 KB, uploaded 31 January 2019]
  |  

Abstract

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
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Supplementary material

SciFeed

Share & Cite This Article

MDPI and ACS Style

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.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top