| Téléchargement | - Voir la version finale : Only a small fraction of cells produce assembled capsids during transfection‐based manufacturing of adeno‐associated virus vectors (PDF, 1.4 Mio)
|
|---|
| DOI | Trouver le DOI : https://doi.org/10.1002/bit.28068 |
|---|
| Auteur | Rechercher : Dash, Shantoshini; Rechercher : Sharon, David M.; Rechercher : Mullick, Alaka1; Rechercher : Kamen, Amine A.1Identifiant ORCID : https://orcid.org/0000-0001-9110-8815 |
|---|
| Affiliation | - Conseil national de recherches du Canada. Thérapeutique en santé humaine
|
|---|
| Bailleur de fonds | Rechercher : National Research Council Canada |
|---|
| Format | Texte, Article |
|---|
| Sujet | adeno‐associated virus; gene therapy; transfection; viral vectors |
|---|
| Résumé | Plasmid transfection of mammalian cells is the dominant platform used to produce adeno-associated virus (AAV) vectors for clinical and research applications. Low yields from this platform currently make it difficult to supply these activities with adequate material. In an effort to better understand the current limitations of transfection-based manufacturing, this study examines what proportion of cells in a model transfection produce appreciable amounts of assembled AAV capsid. Using conformation-specific antibody staining and flow cytometry, we report the surprising result that despite obtaining high transfection efficiencies and nominal vector yields in our model system, only 5%–10% of cells appear to produce measurable levels of assembled AAV capsids. This finding implies that considerable increases in vector titer could be realized through increasing the proportion of productive cells. Furthermore, we suggest that the flow cytometry assay used here to quantify productive cells may be a useful metric for future optimization of transfection-based AAV vector manufacturing platforms. |
|---|
| Date de publication | 2022-02-19 |
|---|
| Maison d’édition | Wiley |
|---|
| Licence | |
|---|
| Dans | |
|---|
| Langue | anglais |
|---|
| Publications évaluées par des pairs | Oui |
|---|
| Exporter la notice | Exporter en format RIS |
|---|
| Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
|---|
| Identificateur de l’enregistrement | 158c7b3e-9b7d-4345-bb09-590669d6ea35 |
|---|
| Enregistrement créé | 2024-01-29 |
|---|
| Enregistrement modifié | 2024-01-29 |
|---|
