Goebel, Sven;
Jaen, Karim EE;
Dorn, Marie;
Neumeyer, Victoria;
Jordan, Ingo;
Sandig, Volker;
Reichl, Udo;
... Genzel, Yvonne; + view all
(2023)
Process intensification strategies toward cell culture-based high-yield production of a fusogenic oncolytic virus.
Biotechnology and Bioengineering
, 120
(9)
pp. 2639-2657.
10.1002/bit.28353.
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Abstract
We present a proof-of-concept study for production of a recombinant vesicular stomatitis virus (rVSV)-based fusogenic oncolytic virus (OV), rVSV-Newcastle disease virus (NDV), at high cell densities (HCD). Based on comprehensive experiments in 1 L stirred tank reactors (STRs) in batch mode, first optimization studies at HCD were carried out in semi-perfusion in small-scale cultivations using shake flasks. Further, a perfusion process was established using an acoustic settler for cell retention. Growth, production yields, and process-related impurities were evaluated for three candidate cell lines (AGE1.CR, BHK-21, HEK293SF)infected at densities ranging from 15 to 30 × 106 cells/mL. The acoustic settler allowed continuous harvesting of rVSV-NDV with high cell retention efficiencies (above 97%) and infectious virus titers (up to 2.4 × 109 TCID50/mL), more than 4–100 times higher than for optimized batch processes. No decrease in cell-specific virus yield (CSVY) was observed at HCD, regardless of the cell substrate. Taking into account the accumulated number of virions both from the harvest and bioreactor, a 15–30 fold increased volumetric virus productivity for AGE1.CR and HEK293SF was obtained compared to batch processes performed at the same scale. In contrast to all previous findings, formation of syncytia was observed at HCD for the suspension cells BHK 21 and HEK293SF. Oncolytic potency was not affected compared to production in batch mode. Overall, our study describes promising options for the establishment of perfusion processes for efficient large-scale manufacturing of fusogenic rVSV-NDV at HCD for all three candidate cell lines.
| Type: | Article |
|---|---|
| Title: | Process intensification strategies toward cell culture-based high-yield production of a fusogenic oncolytic virus |
| Location: | United States |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1002/bit.28353 |
| Publisher version: | https://doi.org/10.1002/bit.28353 |
| Language: | English |
| Additional information: | This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. See: http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| Keywords: | Science & Technology, Life Sciences & Biomedicine, Biotechnology & Applied Microbiology, cell culture-based production, fusogenic oncolytic virus, high cell density culture, perfusion, process intensification, upstream processing, INFLUENZA-VIRUS, DENSITY CULTIVATIONS, SCALABLE PRODUCTION, PRODUCTION KINETICS, ADENOVIRUS VECTOR, SUSPENSION, PERFUSION, INSULIN, LINES, OPTIMIZATION |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10204964 |
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