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

Eduard Ayuso, Véronique Blouin, Martin Lock, Susan Mcgorray, Xavier Leon, Mauricio R. Alvira, Alberto Auricchio, Stephanie Bucher, Abdelwahed Chtarto, K. Reed Clark, Christophe Darmon, Monica Doria, Will Fountain, Guangping Gao, Kai Gao, Mauro Giacca, Juergen Kleinschmidt, Barbara Leuchs, Catherine Melas, Hiroaki MizukamiMarcus Müller, Yvet Noordman, Olivier Bockstael, Keiya Ozawa, Catherine Pythoud, Marina Sumaroka, Richard Surosky, Liliane Tenenbaum, Inge Van Der Linden, Brigitte Weins, J. Fraser Wright, Xinhua Zhang, Lorena Zentilin, Fatima Bosch, Richard O. Snyder, Philippe Moullier

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© Mary Ann Liebert, Inc. 2014. 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, 5.50×1011 pt/ml; CI, 4.26×1011 to 6.75×1011 pt/ml), vector genomes (mean, 5.75×1011 vg/ml; CI, 3.05×1011 to 1.09×1012 vg/ml), and infectious units (mean, 1.26×109 IU/ml; CI, 6.46×108 to 2.51×109 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.
Idioma originalEnglish
Pàgines (de-a)977-987
RevistaHuman Gene Therapy
Estat de la publicacióPublicada - 1 de gen. 2014


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