Cell culture-based production of defective interfering influenza A virus particles in perfusion mode using an alternating tangential flow filtration system.
Alternating tangential flow filtration (ATF)
Antiviral
Bioreactor
Cell culture–based production
Defective interfering particles (DIPs)
Influenza A virus
Perfusion cultivation
Journal
Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612
Informations de publication
Date de publication:
Oct 2021
Oct 2021
Historique:
received:
11
06
2021
accepted:
26
08
2021
revised:
25
08
2021
pubmed:
15
9
2021
medline:
9
10
2021
entrez:
14
9
2021
Statut:
ppublish
Résumé
Respiratory diseases including influenza A virus (IAV) infections represent a major threat to human health. While the development of a vaccine requires a lot of time, a fast countermeasure could be the use of defective interfering particles (DIPs) for antiviral therapy. IAV DIPs are usually characterized by a large internal deletion in one viral RNA segment. Consequentially, DIPs can only propagate in presence of infectious standard viruses (STVs), compensating the missing gene function. Here, they interfere with and suppress the STV replication and might act "universally" against many IAV subtypes. We recently reported a production system for purely clonal DIPs utilizing genetically modified cells. In the present study, we established an automated perfusion process for production of a DIP, called DI244, using an alternating tangential flow filtration (ATF) system for cell retention. Viable cell concentrations and DIP titers more than 10 times higher than for a previously reported batch cultivation were observed. Furthermore, we investigated a novel tubular cell retention device for its potential for continuous virus harvesting into the permeate. Very comparable performances to typically used hollow fiber membranes were found during the cell growth phase. During the virus replication phase, the tubular membrane, in contrast to the hollow fiber membrane, allowed 100% of the produced virus particles to pass through. To our knowledge, this is the first time a continuous virus harvest was shown for a membrane-based perfusion process. Overall, the process established offers interesting possibilities for advanced process integration strategies for next-generation virus particle and virus vector manufacturing.Key points• An automated perfusion process for production of IAV DIPs was established.• DIP titers of 7.40E + 9 plaque forming units per mL were reached.• A novel tubular cell retention device enabled continuous virus harvesting.
Identifiants
pubmed: 34519855
doi: 10.1007/s00253-021-11561-y
pii: 10.1007/s00253-021-11561-y
pmc: PMC8437742
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
7251-7264Subventions
Organisme : Defense Advanced Research Projects Agency
ID : W911NF-17-2-0012
Informations de copyright
© 2021. The Author(s).
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