Optimization of YF17D-Vectored Zika Vaccine Production by Employing Small-Molecule Viral Sensitizers to Enhance Yields

<b>Background:</b> Modern viral vector production needs to consider process intensification for higher yields from smaller production volumes. However, innate antiviral immunity triggered in the producer cell may limit virus replication. While commonly used cell lines (e.g., Vero or E1A-...

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Main Authors: Sven Göbel, Tilia Zinnecker, Ingo Jordan, Volker Sandig, Andrea Vervoort, Jondavid de Jong, Jean-Simon Diallo, Peter Satzer, Manfred Satzer, Kai Dallmeier, Udo Reichl, Yvonne Genzel
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Language:English
Published: MDPI AG 2025-07-01
Series:Vaccines
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Online Access:https://www.mdpi.com/2076-393X/13/7/757
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author Sven Göbel
Tilia Zinnecker
Ingo Jordan
Volker Sandig
Andrea Vervoort
Jondavid de Jong
Jean-Simon Diallo
Peter Satzer
Manfred Satzer
Kai Dallmeier
Udo Reichl
Yvonne Genzel
author_facet Sven Göbel
Tilia Zinnecker
Ingo Jordan
Volker Sandig
Andrea Vervoort
Jondavid de Jong
Jean-Simon Diallo
Peter Satzer
Manfred Satzer
Kai Dallmeier
Udo Reichl
Yvonne Genzel
author_sort Sven Göbel
collection DOAJ
description <b>Background:</b> Modern viral vector production needs to consider process intensification for higher yields from smaller production volumes. However, innate antiviral immunity triggered in the producer cell may limit virus replication. While commonly used cell lines (e.g., Vero or E1A-immortalised cells) are already compromised in antiviral pathways, the redundancy of innate signaling complicates host cell optimization by genetic engineering. Small molecules that are hypothesized to target antiviral pathways (Viral Sensitizers, VSEs) added to the culture media offer a versatile alternative to genetic modifications to increase permissiveness and, thus, viral yields across multiple cell lines. <b>Methods:</b> To explore how the yield for a chimeric Zika vaccine candidate (YF-ZIK) could be further be increased in an intensified bioprocess, we used spin tubes or an Ambr15 high-throughput microbioreactor system as scale-down models to optimize the dosing for eight VSEs in three host cell lines (AGE1.CR.pIX, BHK-21, and HEK293-F) based on their tolerability. <b>Results:</b> Addition of VSEs to an already optimized infection process significantly increased infectious titers by up to sevenfold for all three cell lines tested. The development of multi-component VSE formulations using a design of experiments approach allowed further synergistic titer increases in AGE1.CR.pIX cells. Scale-up to 1 L stirred-tank bioreactors and 3D-printed mimics of 200 or 2000 L reactors resulted in up to threefold and eightfold increases, respectively. <b>Conclusions:</b> Addition of single VSEs or combinations thereof allowed a further increase in YF-ZIK titers beyond the yield of an already optimized, highly intensified process. The described approach validates the use of VSEs and can be instructive for optimizing other virus production processes.
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spelling doaj-art-657b23d4714c4deeb69d39cbcfe6951f2025-08-20T03:32:16ZengMDPI AGVaccines2076-393X2025-07-0113775710.3390/vaccines13070757Optimization of YF17D-Vectored Zika Vaccine Production by Employing Small-Molecule Viral Sensitizers to Enhance YieldsSven Göbel0Tilia Zinnecker1Ingo Jordan2Volker Sandig3Andrea Vervoort4Jondavid de Jong5Jean-Simon Diallo6Peter Satzer7Manfred Satzer8Kai Dallmeier9Udo Reichl10Yvonne Genzel11Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106 Magdeburg, GermanyBioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106 Magdeburg, GermanyProBioGen AG, 13086 Berlin, GermanyProBioGen AG, 13086 Berlin, GermanyVirica Biotech, Ottawa, ON K1Y 2C5, CanadaVirica Biotech, Ottawa, ON K1Y 2C5, CanadaVirica Biotech, Ottawa, ON K1Y 2C5, Canadap4b GmbH, 2120 Wolkersdorf, Austriap4b GmbH, 2120 Wolkersdorf, AustriaKU Leuven Department of Microbiology, Immunology & Transplantation, Rega Institute, Molecular Vaccinology and Vaccine Discovery (MVVD), 3000 Leuven, BelgiumBioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106 Magdeburg, GermanyBioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106 Magdeburg, Germany<b>Background:</b> Modern viral vector production needs to consider process intensification for higher yields from smaller production volumes. However, innate antiviral immunity triggered in the producer cell may limit virus replication. While commonly used cell lines (e.g., Vero or E1A-immortalised cells) are already compromised in antiviral pathways, the redundancy of innate signaling complicates host cell optimization by genetic engineering. Small molecules that are hypothesized to target antiviral pathways (Viral Sensitizers, VSEs) added to the culture media offer a versatile alternative to genetic modifications to increase permissiveness and, thus, viral yields across multiple cell lines. <b>Methods:</b> To explore how the yield for a chimeric Zika vaccine candidate (YF-ZIK) could be further be increased in an intensified bioprocess, we used spin tubes or an Ambr15 high-throughput microbioreactor system as scale-down models to optimize the dosing for eight VSEs in three host cell lines (AGE1.CR.pIX, BHK-21, and HEK293-F) based on their tolerability. <b>Results:</b> Addition of VSEs to an already optimized infection process significantly increased infectious titers by up to sevenfold for all three cell lines tested. The development of multi-component VSE formulations using a design of experiments approach allowed further synergistic titer increases in AGE1.CR.pIX cells. Scale-up to 1 L stirred-tank bioreactors and 3D-printed mimics of 200 or 2000 L reactors resulted in up to threefold and eightfold increases, respectively. <b>Conclusions:</b> Addition of single VSEs or combinations thereof allowed a further increase in YF-ZIK titers beyond the yield of an already optimized, highly intensified process. The described approach validates the use of VSEs and can be instructive for optimizing other virus production processes.https://www.mdpi.com/2076-393X/13/7/757bioprocess engineeringvectored live-attenuated vaccinesviral sensitizerssmall moleculesdesign of experimentsantiviral defense
spellingShingle Sven Göbel
Tilia Zinnecker
Ingo Jordan
Volker Sandig
Andrea Vervoort
Jondavid de Jong
Jean-Simon Diallo
Peter Satzer
Manfred Satzer
Kai Dallmeier
Udo Reichl
Yvonne Genzel
Optimization of YF17D-Vectored Zika Vaccine Production by Employing Small-Molecule Viral Sensitizers to Enhance Yields
Vaccines
bioprocess engineering
vectored live-attenuated vaccines
viral sensitizers
small molecules
design of experiments
antiviral defense
title Optimization of YF17D-Vectored Zika Vaccine Production by Employing Small-Molecule Viral Sensitizers to Enhance Yields
title_full Optimization of YF17D-Vectored Zika Vaccine Production by Employing Small-Molecule Viral Sensitizers to Enhance Yields
title_fullStr Optimization of YF17D-Vectored Zika Vaccine Production by Employing Small-Molecule Viral Sensitizers to Enhance Yields
title_full_unstemmed Optimization of YF17D-Vectored Zika Vaccine Production by Employing Small-Molecule Viral Sensitizers to Enhance Yields
title_short Optimization of YF17D-Vectored Zika Vaccine Production by Employing Small-Molecule Viral Sensitizers to Enhance Yields
title_sort optimization of yf17d vectored zika vaccine production by employing small molecule viral sensitizers to enhance yields
topic bioprocess engineering
vectored live-attenuated vaccines
viral sensitizers
small molecules
design of experiments
antiviral defense
url https://www.mdpi.com/2076-393X/13/7/757
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