A rapid Focus-Forming Assay for quantification of infectious adenoviral vectors.
Adenovirus
Focus-Forming Assay
Plaque assay
TCID(50) assay
Journal
Journal of virological methods
ISSN: 1879-0984
Titre abrégé: J Virol Methods
Pays: Netherlands
ID NLM: 8005839
Informations de publication
Date de publication:
11 2021
11 2021
Historique:
received:
27
04
2021
revised:
09
08
2021
accepted:
20
08
2021
pubmed:
27
8
2021
medline:
22
3
2022
entrez:
26
8
2021
Statut:
ppublish
Résumé
Currently available methods to titrate adenoviral vectors (AdV) in the absence of a gene reporter such as GFP, are either time-consuming or not very reproducible. A Focus-Forming Assay (FFA) for quantification of infectious AdV particles followed by automated focus counting was developed using new monoclonal antibodies (mAbs) against the human adenovirus type 5. Briefly, in this method, 96-well plates of HEK293A cells were infected with 2-fold dilutions of AdV at seeding time. Forty eight hours post-infection, the cells were fixed with methanol. The cells were then incubated with each mAb followed by a FITC conjugated anti-mouse antibody. The plates were scanned and positive cells counted using an automated fluorescence microscopy system. The results of the FFA were compared with the plaque assay and the TCID
Identifiants
pubmed: 34437873
pii: S0166-0934(21)00206-8
doi: 10.1016/j.jviromet.2021.114267
pii:
doi:
Substances chimiques
Antibodies, Monoclonal
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
114267Informations de copyright
Crown Copyright © 2021. Published by Elsevier B.V. All rights reserved.