Manufacturing and characterization of a recombinant adeno-associated virus type 8 reference standard material.

Détails

Ressource 1Télécharger: BIB_3BB1F3A8DF53.P001.pdf (2655.16 [Ko])
Etat: Public
Version: Author's accepted manuscript
ID Serval
serval:BIB_3BB1F3A8DF53
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Manufacturing and characterization of a recombinant adeno-associated virus type 8 reference standard material.
Périodique
Human Gene Therapy
Auteur(s)
Ayuso E., Blouin V., Lock M., McGorray S., Leon X., Alvira M.R., Auricchio A., Bucher S., Chtarto A., Clark K.R., Darmon C., Doria M., Fountain W., Gao G., Gao K., Giacca M., Kleinschmidt J., Leuchs B., Melas C., Mizukami H., Müller M., Noordman Y., Bockstael O., Ozawa K., Pythoud C., Sumaroka M., Surosky R., Tenenbaum L., van der Linden I., Weins B., Wright J.F., Zhang X., Zentilin L., Bosch F., Snyder R.O., Moullier P.
ISSN
1557-7422 (Electronic)
ISSN-L
1043-0342
Statut éditorial
Publié
Date de publication
2014
Peer-reviewed
Oui
Volume
25
Numéro
11
Pages
977-987
Langue
anglais
Notes
Publication types: Journal Article
Résumé
Gene therapy approaches using recombinant adeno-associated virus serotype 2 (rAAV2) and serotype 8 (rAAV8) have achieved significant clinical benefits. The generation of rAAV Reference Standard Materials (RSM) is key to providing points of reference for particle titer, vector genome titer, and infectious titer for gene transfer vectors. Following the example of the rAAV2RSM, here we have generated and characterized a novel RSM based on rAAV serotype 8. The rAAV8RSM was produced using transient transfection, and the purification was based on density gradient ultracentrifugation. The rAAV8RSM was distributed for characterization along with standard assay protocols to 16 laboratories worldwide. Mean titers and 95% confidence intervals were determined for capsid particles (mean, 5.50×10(11) pt/ml; CI, 4.26×10(11) to 6.75×10(11) pt/ml), vector genomes (mean, 5.75×10(11) vg/ml; CI, 3.05×10(11) to 1.09×10(12) vg/ml), and infectious units (mean, 1.26×10(9) IU/ml; CI, 6.46×10(8) to 2.51×10(9) IU/ml). Notably, there was a significant degree of variation between institutions for each assay despite the relatively tight correlation of assay results within an institution. This outcome emphasizes the need to use RSMs to calibrate the titers of rAAV vectors in preclinical and clinical studies at a time when the field is maturing rapidly. The rAAV8RSM has been deposited at the American Type Culture Collection (VR-1816) and is available to the scientific community.
Pubmed
Web of science
Création de la notice
27/12/2014 10:31
Dernière modification de la notice
20/08/2019 14:31
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