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Exploring nutrient supplementation and bioprocess optimization to improve the production of lentiviral vectors in serum-free medium suspension cultures

2024-01Artigo científico Acesso aberto
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dc.contributor.authorVaz, Tiago A.
dc.contributor.authorRodrigues, Ana F.
dc.contributor.authorCoroadinha, Ana S.
dc.contributor.institutionInstituto de Tecnologia Química e Biológica António Xavier (ITQB)
dc.contributor.pblWiley
dc.date.accessioned2024-10-01T22:23:42Z
dc.date.available2024-10-01T22:23:42Z
dc.date.issued2024-01
dc.descriptionFunding Information: T.A.V. acknowledges FCT for the award of the individual grant SFRH/BD/145270/2019. This work was funded by Fundação para a Ciência e Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through national funds to iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). Publisher Copyright: © 2023 The Authors. Biotechnology Journal published by Wiley-VCH GmbH.
dc.description.abstractThe use of lentiviral vectors (LV) in gene therapy has been growing in recent years. To meet the increasing clinical demand, LV production platforms will benefit from improved productivity and scalability to enable cost-effective manufacture of LV-based therapies. Here we report the adaptation of 293T cells to serum-free suspension cultures and the improvement of LV yields through transfection parameters optimization, process intensification and medium supplementation with nutrient boosters. Cells were sequentially adapted to different serum-free culture media, transfection parameters were optimized and the two best-performing conditions were selected to explore process intensification by increasing cell density at the time of transfection. LV production at higher cell densities increased volumetric titers up to 12-fold and lipid supplementation was the most efficient metabolic optimization strategy further enhancing LV productivity by 3-fold. Furthermore, cell concentration was identified and validated as an important source of transfection variability impairing cellular uptake of DNA polyplexes, impacting transfection efficiency and reducing LV titers down to 6-fold. This work contributes to improving LV-based gene therapy by establishing new scalable manufacturing platforms and providing key metabolic insights, unveiling important bioreaction parameters to improve vector yields.en
dc.description.versionpublishersversion
dc.description.versionpublished
dc.format.extent1072984
dc.identifier.doi10.1002/biot.202300212
dc.identifier.issn1860-6768
dc.identifier.otherPURE: 90689609
dc.identifier.otherPURE UUID: 5728a087-38d8-4ed9-b7c3-8381dee61868
dc.identifier.otherScopus: 85164213504
dc.identifier.otherPubMed: 37903159
dc.identifier.urihttp://hdl.handle.net/10362/172811
dc.identifier.urlhttps://www.scopus.com/pages/publications/85164213504
dc.language.isoeng
dc.peerreviewedyes
dc.subjectbioprocess engineering
dc.subjectgene therapy
dc.subjectlentiviral vectors
dc.subjectserum-free medium suspension culture
dc.subjecttransient transfection
dc.subjectApplied Microbiology and Biotechnology
dc.subjectMolecular Medicine
dc.titleExploring nutrient supplementation and bioprocess optimization to improve the production of lentiviral vectors in serum-free medium suspension culturesen
dc.typejournal article
degois.publication.issue1
degois.publication.titleBiotechnology Journal
degois.publication.volume19
dspace.entity.typePublication
rcaap.rightsopenAccess

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