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High titer MVA and influenza A virus production using a hybrid fed-batch/perfusion strategy with an ATF system
A cultivation strategy to increase the productivity of Modified Vaccinia Ankara (MVA) virus in high-cell density processes is presented. Based on an approach developed in shake flask cultures, this strategy was established in benchtop bioreactors, comprising the growth of suspension AGE1.CR.pIX cell...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6447503/ https://www.ncbi.nlm.nih.gov/pubmed/30796494 http://dx.doi.org/10.1007/s00253-019-09694-2 |
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author | Vázquez-Ramírez, Daniel Jordan, Ingo Sandig, Volker Genzel, Yvonne Reichl, Udo |
author_facet | Vázquez-Ramírez, Daniel Jordan, Ingo Sandig, Volker Genzel, Yvonne Reichl, Udo |
author_sort | Vázquez-Ramírez, Daniel |
collection | PubMed |
description | A cultivation strategy to increase the productivity of Modified Vaccinia Ankara (MVA) virus in high-cell density processes is presented. Based on an approach developed in shake flask cultures, this strategy was established in benchtop bioreactors, comprising the growth of suspension AGE1.CR.pIX cells to high cell densities in a chemically defined medium before infection with the MVA-CR19 virus strain. First, a perfusion regime was established to optimize the cell growth phase. Second, a fed-batch regime was chosen for the initial infection phase to facilitate virus uptake and cell-to-cell spreading. Afterwards, a switch to perfusion enabled the continuous supply of nutrients for the late stages of virus propagation. With maximum infectious titers of 1.0 × 10(10) IU/mL, this hybrid fed-batch/perfusion strategy increased product titers by almost one order of magnitude compared to conventional batch cultivations. Finally, this strategy was also applied to the production of influenza A/PR/8/34 (H1N1) virus considered for manufacturing of inactivated vaccines. Using the same culture system, a total number of 3.8 × 10(10) virions/mL was achieved. Overall, comparable or even higher cell-specific virus yields and volumetric productivities were obtained using the same cultivation systems as for the conventional batch cultivations. In addition, most viral particles were found in the culture supernatant, which can simplify further downstream operations, in particular for MVA viruses. Considering the current availability of well-described perfusion/cell retention technologies, the present strategy may contribute to the development of new approaches for viral vaccine production. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-019-09694-2) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6447503 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-64475032019-04-17 High titer MVA and influenza A virus production using a hybrid fed-batch/perfusion strategy with an ATF system Vázquez-Ramírez, Daniel Jordan, Ingo Sandig, Volker Genzel, Yvonne Reichl, Udo Appl Microbiol Biotechnol Biotechnological Products and Process Engineering A cultivation strategy to increase the productivity of Modified Vaccinia Ankara (MVA) virus in high-cell density processes is presented. Based on an approach developed in shake flask cultures, this strategy was established in benchtop bioreactors, comprising the growth of suspension AGE1.CR.pIX cells to high cell densities in a chemically defined medium before infection with the MVA-CR19 virus strain. First, a perfusion regime was established to optimize the cell growth phase. Second, a fed-batch regime was chosen for the initial infection phase to facilitate virus uptake and cell-to-cell spreading. Afterwards, a switch to perfusion enabled the continuous supply of nutrients for the late stages of virus propagation. With maximum infectious titers of 1.0 × 10(10) IU/mL, this hybrid fed-batch/perfusion strategy increased product titers by almost one order of magnitude compared to conventional batch cultivations. Finally, this strategy was also applied to the production of influenza A/PR/8/34 (H1N1) virus considered for manufacturing of inactivated vaccines. Using the same culture system, a total number of 3.8 × 10(10) virions/mL was achieved. Overall, comparable or even higher cell-specific virus yields and volumetric productivities were obtained using the same cultivation systems as for the conventional batch cultivations. In addition, most viral particles were found in the culture supernatant, which can simplify further downstream operations, in particular for MVA viruses. Considering the current availability of well-described perfusion/cell retention technologies, the present strategy may contribute to the development of new approaches for viral vaccine production. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-019-09694-2) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2019-02-23 2019 /pmc/articles/PMC6447503/ /pubmed/30796494 http://dx.doi.org/10.1007/s00253-019-09694-2 Text en © The Author(s) 2019 OpenAccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Biotechnological Products and Process Engineering Vázquez-Ramírez, Daniel Jordan, Ingo Sandig, Volker Genzel, Yvonne Reichl, Udo High titer MVA and influenza A virus production using a hybrid fed-batch/perfusion strategy with an ATF system |
title | High titer MVA and influenza A virus production using a hybrid fed-batch/perfusion strategy with an ATF system |
title_full | High titer MVA and influenza A virus production using a hybrid fed-batch/perfusion strategy with an ATF system |
title_fullStr | High titer MVA and influenza A virus production using a hybrid fed-batch/perfusion strategy with an ATF system |
title_full_unstemmed | High titer MVA and influenza A virus production using a hybrid fed-batch/perfusion strategy with an ATF system |
title_short | High titer MVA and influenza A virus production using a hybrid fed-batch/perfusion strategy with an ATF system |
title_sort | high titer mva and influenza a virus production using a hybrid fed-batch/perfusion strategy with an atf system |
topic | Biotechnological Products and Process Engineering |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6447503/ https://www.ncbi.nlm.nih.gov/pubmed/30796494 http://dx.doi.org/10.1007/s00253-019-09694-2 |
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