Process Development for Newcastle Disease Virus-Vectored Vaccines in Serum-Free Vero Cell Suspension Cultures

The ongoing COVID-19 pandemic drew global attention to infectious diseases, attracting numerous resources for development of pandemic preparedness plans and vaccine platforms—technologies with robust manufacturing processes that can quickly be pivoted to target emerging diseases. Newcastle Disease V...

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Autores principales:	Julia Puppin Chaves Fulber, Omar Farnós, Sascha Kiesslich, Zeyu Yang, Shantoshini Dash, Leonardo Susta, Sarah K. Wootton, Amine A. Kamen
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	Newcastle Disease Virus Vero suspension culture viral vaccine bioprocess bioreactor production vaccine production platform COVID-19 Medicine R
Acceso en línea:	https://doaj.org/article/b7aba1817a4c4c60b48596beb30861ff
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spelling	oai:doaj.org-article:b7aba1817a4c4c60b48596beb30861ff2021-11-25T19:11:24ZProcess Development for Newcastle Disease Virus-Vectored Vaccines in Serum-Free Vero Cell Suspension Cultures10.3390/vaccines91113352076-393Xhttps://doaj.org/article/b7aba1817a4c4c60b48596beb30861ff2021-11-01T00:00:00Zhttps://www.mdpi.com/2076-393X/9/11/1335https://doaj.org/toc/2076-393XThe ongoing COVID-19 pandemic drew global attention to infectious diseases, attracting numerous resources for development of pandemic preparedness plans and vaccine platforms—technologies with robust manufacturing processes that can quickly be pivoted to target emerging diseases. Newcastle Disease Virus (NDV) has been studied as a viral vector for human and veterinary vaccines, but its production relies heavily on embryonated chicken eggs, with very few studies producing NDV in cell culture. Here, NDV is produced in suspension Vero cells, and analytical assays (TCID<sub>50</sub> and ddPCR) are developed to quantify infectious and total viral titer. NDV-GFP and NDV-FLS (SARS-CoV-2 full-length spike protein) constructs were adapted to replicate in Vero and HEK293 suspension cultures using serum-free media, while fine-tuning parameters such as MOI, temperature, and trypsin concentration. Shake flask productions with Vero cells resulted in infectious titers of 1.07 × 10<sup>8</sup> TCID<sub>50</sub>/mL for NDV-GFP and 1.33 × 10<sup>8</sup> TCID<sub>50</sub>/mL for NDV-FLS. Production in 1 L batch bioreactors also resulted in high titers in culture supernatants, reaching 2.37 × 10<sup>8</sup> TCID<sub>50</sub>/mL for NDV-GFP and 3.16 × 10<sup>7</sup> TCID<sub>50</sub>/mL for NDV-FLS. This shows effective NDV production in cell culture, building the basis for a scalable vectored-vaccine manufacturing process that can be applied to different targets.Julia Puppin Chaves FulberOmar FarnósSascha KiesslichZeyu YangShantoshini DashLeonardo SustaSarah K. WoottonAmine A. KamenMDPI AGarticleNewcastle Disease VirusVero suspension cultureviral vaccine bioprocessbioreactor productionvaccine production platformCOVID-19MedicineRENVaccines, Vol 9, Iss 1335, p 1335 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Newcastle Disease Virus Vero suspension culture viral vaccine bioprocess bioreactor production vaccine production platform COVID-19 Medicine R
spellingShingle	Newcastle Disease Virus Vero suspension culture viral vaccine bioprocess bioreactor production vaccine production platform COVID-19 Medicine R Julia Puppin Chaves Fulber Omar Farnós Sascha Kiesslich Zeyu Yang Shantoshini Dash Leonardo Susta Sarah K. Wootton Amine A. Kamen Process Development for Newcastle Disease Virus-Vectored Vaccines in Serum-Free Vero Cell Suspension Cultures
description	The ongoing COVID-19 pandemic drew global attention to infectious diseases, attracting numerous resources for development of pandemic preparedness plans and vaccine platforms—technologies with robust manufacturing processes that can quickly be pivoted to target emerging diseases. Newcastle Disease Virus (NDV) has been studied as a viral vector for human and veterinary vaccines, but its production relies heavily on embryonated chicken eggs, with very few studies producing NDV in cell culture. Here, NDV is produced in suspension Vero cells, and analytical assays (TCID<sub>50</sub> and ddPCR) are developed to quantify infectious and total viral titer. NDV-GFP and NDV-FLS (SARS-CoV-2 full-length spike protein) constructs were adapted to replicate in Vero and HEK293 suspension cultures using serum-free media, while fine-tuning parameters such as MOI, temperature, and trypsin concentration. Shake flask productions with Vero cells resulted in infectious titers of 1.07 × 10<sup>8</sup> TCID<sub>50</sub>/mL for NDV-GFP and 1.33 × 10<sup>8</sup> TCID<sub>50</sub>/mL for NDV-FLS. Production in 1 L batch bioreactors also resulted in high titers in culture supernatants, reaching 2.37 × 10<sup>8</sup> TCID<sub>50</sub>/mL for NDV-GFP and 3.16 × 10<sup>7</sup> TCID<sub>50</sub>/mL for NDV-FLS. This shows effective NDV production in cell culture, building the basis for a scalable vectored-vaccine manufacturing process that can be applied to different targets.
format	article
author	Julia Puppin Chaves Fulber Omar Farnós Sascha Kiesslich Zeyu Yang Shantoshini Dash Leonardo Susta Sarah K. Wootton Amine A. Kamen
author_facet	Julia Puppin Chaves Fulber Omar Farnós Sascha Kiesslich Zeyu Yang Shantoshini Dash Leonardo Susta Sarah K. Wootton Amine A. Kamen
author_sort	Julia Puppin Chaves Fulber
title	Process Development for Newcastle Disease Virus-Vectored Vaccines in Serum-Free Vero Cell Suspension Cultures
title_short	Process Development for Newcastle Disease Virus-Vectored Vaccines in Serum-Free Vero Cell Suspension Cultures
title_full	Process Development for Newcastle Disease Virus-Vectored Vaccines in Serum-Free Vero Cell Suspension Cultures
title_fullStr	Process Development for Newcastle Disease Virus-Vectored Vaccines in Serum-Free Vero Cell Suspension Cultures
title_full_unstemmed	Process Development for Newcastle Disease Virus-Vectored Vaccines in Serum-Free Vero Cell Suspension Cultures
title_sort	process development for newcastle disease virus-vectored vaccines in serum-free vero cell suspension cultures
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/b7aba1817a4c4c60b48596beb30861ff
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Process Development for Newcastle Disease Virus-Vectored Vaccines in Serum-Free Vero Cell Suspension Cultures

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