High Cell Density Perfusion Process of Quail Cells Producing Oncolytic rVSV‐NDV
ABSTRACT Oncolytic viruses as agents for the treatment of various types of cancer have demonstrated their potential in many clinical studies over the past decades. In particular, rVSV‐NDV (a recombinant vesicular stomatitis virus [VSV] construct with fusogenic Newcastle disease virus glycoproteins)...
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Wiley-VCH
2025-07-01
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| Series: | Engineering in Life Sciences |
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| Online Access: | https://doi.org/10.1002/elsc.70035 |
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| author | Lennart Jacobtorweihe Sven Göbel Markus Wolschek Jennifer Altomonte Udo Reichl Yvonne Genzel |
| author_facet | Lennart Jacobtorweihe Sven Göbel Markus Wolschek Jennifer Altomonte Udo Reichl Yvonne Genzel |
| author_sort | Lennart Jacobtorweihe |
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| description | ABSTRACT Oncolytic viruses as agents for the treatment of various types of cancer have demonstrated their potential in many clinical studies over the past decades. In particular, rVSV‐NDV (a recombinant vesicular stomatitis virus [VSV] construct with fusogenic Newcastle disease virus glycoproteins) shows promising preclinical results. This is due to its safety profile, immunostimulatory effects, and efficacy based on strong syncytia formation. Since virotherapy requires a high input of infectious viruses, efficient production processes are needed. Good manufacturing practice (GMP)‐compliant CCX.E10 cells have been previously reported as a high‐titer‐producing rVSV‐NDV candidate in batch mode. Here, semi‐perfusion was used to test quail‐originated CCX.E10 cells for rVSV‐NDV production at high cell densities and in different cell culture media. The best condition was transferred to a full perfusion process in a 3 L bioreactor using a tangential follow depth filtration (TFDF) device for cell retention. The integrated depth filter with a pore size of 2–5 µm allowed 99.9% cell retention at viable cell concentrations (VCCs) of up to 20.6 × 106 cells/mL and continuous virus harvesting. With this setup, we were able to produce 1.33 × 109 TCID50/mL infectious virus with a 5‐fold increase in space‐time yield (STY) compared to a batch process as a control. Practical application: Despite significant progress in oncolytic virus development, early research primarily focuses on viral design and therapeutic potential, often overlooking production challenges until later stages. This gap hinders clinical translation, as manufacturing high oncolytic virus doses (up to 10¹¹ infectious particles per injection) remains a major bottleneck. Implementing GMP‐compliant cell substrates alongside perfusion cultures is essential to overcoming the low yields of traditional batch production. These advancements have far‐reaching implications for reducing costs, increasing dose availability, and accelerating the clinical adoption of this promising immunotherapy. |
| format | Article |
| id | doaj-art-b04fea215a1548ebb2d638a8fc44ca30 |
| institution | DOAJ |
| issn | 1618-0240 1618-2863 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Wiley-VCH |
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| series | Engineering in Life Sciences |
| spelling | doaj-art-b04fea215a1548ebb2d638a8fc44ca302025-08-20T02:46:13ZengWiley-VCHEngineering in Life Sciences1618-02401618-28632025-07-01257n/an/a10.1002/elsc.70035High Cell Density Perfusion Process of Quail Cells Producing Oncolytic rVSV‐NDVLennart Jacobtorweihe0Sven Göbel1Markus Wolschek2Jennifer Altomonte3Udo Reichl4Yvonne Genzel5Bioprocess Engineering Max Planck Institute For Dynamics of Complex Technical Systems Magdeburg GermanyBioprocess Engineering Max Planck Institute For Dynamics of Complex Technical Systems Magdeburg GermanyNuvonis Technologies GmbH Vienna AustriaDepartment of Internal Medicine II, Klinikum Rechts Der Isar Technische Universität München Munich GermanyBioprocess Engineering Max Planck Institute For Dynamics of Complex Technical Systems Magdeburg GermanyBioprocess Engineering Max Planck Institute For Dynamics of Complex Technical Systems Magdeburg GermanyABSTRACT Oncolytic viruses as agents for the treatment of various types of cancer have demonstrated their potential in many clinical studies over the past decades. In particular, rVSV‐NDV (a recombinant vesicular stomatitis virus [VSV] construct with fusogenic Newcastle disease virus glycoproteins) shows promising preclinical results. This is due to its safety profile, immunostimulatory effects, and efficacy based on strong syncytia formation. Since virotherapy requires a high input of infectious viruses, efficient production processes are needed. Good manufacturing practice (GMP)‐compliant CCX.E10 cells have been previously reported as a high‐titer‐producing rVSV‐NDV candidate in batch mode. Here, semi‐perfusion was used to test quail‐originated CCX.E10 cells for rVSV‐NDV production at high cell densities and in different cell culture media. The best condition was transferred to a full perfusion process in a 3 L bioreactor using a tangential follow depth filtration (TFDF) device for cell retention. The integrated depth filter with a pore size of 2–5 µm allowed 99.9% cell retention at viable cell concentrations (VCCs) of up to 20.6 × 106 cells/mL and continuous virus harvesting. With this setup, we were able to produce 1.33 × 109 TCID50/mL infectious virus with a 5‐fold increase in space‐time yield (STY) compared to a batch process as a control. Practical application: Despite significant progress in oncolytic virus development, early research primarily focuses on viral design and therapeutic potential, often overlooking production challenges until later stages. This gap hinders clinical translation, as manufacturing high oncolytic virus doses (up to 10¹¹ infectious particles per injection) remains a major bottleneck. Implementing GMP‐compliant cell substrates alongside perfusion cultures is essential to overcoming the low yields of traditional batch production. These advancements have far‐reaching implications for reducing costs, increasing dose availability, and accelerating the clinical adoption of this promising immunotherapy.https://doi.org/10.1002/elsc.70035oncolytic virus productionperfusion processprocess intensificationtangential flow depth filtration |
| spellingShingle | Lennart Jacobtorweihe Sven Göbel Markus Wolschek Jennifer Altomonte Udo Reichl Yvonne Genzel High Cell Density Perfusion Process of Quail Cells Producing Oncolytic rVSV‐NDV Engineering in Life Sciences oncolytic virus production perfusion process process intensification tangential flow depth filtration |
| title | High Cell Density Perfusion Process of Quail Cells Producing Oncolytic rVSV‐NDV |
| title_full | High Cell Density Perfusion Process of Quail Cells Producing Oncolytic rVSV‐NDV |
| title_fullStr | High Cell Density Perfusion Process of Quail Cells Producing Oncolytic rVSV‐NDV |
| title_full_unstemmed | High Cell Density Perfusion Process of Quail Cells Producing Oncolytic rVSV‐NDV |
| title_short | High Cell Density Perfusion Process of Quail Cells Producing Oncolytic rVSV‐NDV |
| title_sort | high cell density perfusion process of quail cells producing oncolytic rvsv ndv |
| topic | oncolytic virus production perfusion process process intensification tangential flow depth filtration |
| url | https://doi.org/10.1002/elsc.70035 |
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