Process intensification of EB66® cell cultivations leads to high-yield yellow fever and Zika virus production

Autor:	Udo Reichl, Yvonne Genzel, Arnaud Leon, Alexander Nikolay, Klaus Schwamborn
Rok vydání:	2018
Předmět:	0301 basic medicine Virus Cultivation viruses Cell Culture Techniques Applied Microbiology and Biotechnology Virus Zika virus Cell Line Capacitance probe 03 medical and health sciences EB66® 0302 clinical medicine Bioreactors Immunity medicine Animals 030212 general & internal medicine biology Flavivirus Yellow fever Viral Vaccines General Medicine Zika Virus biology.organism_classification medicine.disease Virology Biotechnological Products and Process Engineering Perfusion Titer Chemically defined medium 030104 developmental biology Ducks Cell culture Yellow fever virus Biotechnology
Zdroj:	Applied Microbiology and Biotechnology
ISSN:	1432-0614
Popis:	A live-attenuated, human vaccine against mosquito-borne yellow fever virus has been available since the 1930s. The vaccine provides long-lasting immunity and consistent mass vaccination campaigns counter viral spread. However, traditional egg-based vaccine manufacturing requires about 12 months and vaccine supplies are chronically close to shortages. In particular, for urban outbreaks, vaccine demand can be covered rarely by global stockpiling. Thus, there is an urgent need for an improved vaccine production platform, ideally transferable to other flaviviruses including Zika virus. Here, we present a proof-of-concept study regarding cell culture-based yellow fever virus 17D (YFV) and wild-type Zika virus (ZIKV) production using duck embryo-derived EB66® cells. Based on comprehensive studies in shake flasks, 1-L bioreactor systems were operated with scalable hollow fiber-based tangential flow filtration (TFF) and alternating tangential flow filtration (ATF) perfusion systems for process intensification. EB66® cells grew in chemically defined medium to cell concentrations of 1.6 × 108 cells/mL. Infection studies with EB66®-adapted virus led to maximum YFV titers of 7.3 × 108 PFU/mL, which corresponds to about 10 million vaccine doses for the bioreactor harvest. For ZIKV, titers of 1.0 × 1010 PFU/mL were achieved. Processes were automated successfully using a capacitance probe to control perfusion rates based on on-line measured cell concentrations. The use of cryo-bags for direct inoculation of production bioreactors facilitates pre-culture preparation contributing to improved process robustness. In conclusion, this platform is a powerful option for next generation cell culture-based flavivirus vaccine manufacturing. Electronic supplementary material The online version of this article (10.1007/s00253-018-9275-z) contains supplementary material, which is available to authorized users.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::120f830a12e1379f29540589b55bcbe9 https://pubmed.ncbi.nlm.nih.gov/30091043 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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