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Stability of the adeno-associated virus 8 reference standard material

Gene Therapy volume 26, pages 211–215 (2019)Cite this article

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Abstract

Adeno-associated virus (AAV) vectors are extensively used for gene therapy clinical trials. Accurate and standardized titration methods are essential for characterizing and dosing AAV-based drugs and thus to assess their safety and efficacy. To this end, the Reference Standard Materials (RSM) working group generated standards for AAV serotype 2 and serotype 8. The AAV8RSM (ATCC® VR-1816™) was deposited to the American Type Culture Collection in 2014 and is available to the scientific community. Here, three independent laboratories of the RSM working group provide stability data of the AAV8RSM 2 years after the initial characterization and after container relabeling performed at the ATCC. The AAV8RSM showed constant titers across experimental conditions: 1.48 ± 0.62 × 1012 vector genome (vg)/ml, 9.38 ± 11.4 × 108 infectious units (IU)/ml and 5.76 ± 2.39 × 1011 total particles (p)/ml as determined by qPCR, TCID50 and ELISA, respectively. Additionally, the AAV8RSM capsid protein integrity assessed by SDS-PAGE was equivalent to the original analyses. In conclusion, the AAV8RSM titers remained stable for two years under appropriate storage conditions ( <−70° C). The use of RSM is strongly recommended and endorsed by regulatory agencies to normalize laboratory internal controls and to provide accurate titration of AAV vectors lots.

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Acknowledgements

We thank ATCC for their contribution labeling the vials and sending materials to the testing laboratories involved in this study. We thank Sven Kuhlendahl and Progen team for providing data, helpful discussions and the ELISA kits used in this study along with Diego Matayoshi, Kristy Guo, Jessica Stone and Kathy Lastra at Brammer Bio. Financial support to E.A.’s laboratory was provided by INSERM, University of Nantes, CHU of Nantes, Fondation d’Entreprises pour la Thérapie Génique en Pays de la Loire, and Région Pays de la Loire and Pre-industrial gene therapy consortium (Agence Nationale de la Recherche - Investissements d’Avenir, grant number 10-DPBS-0001).

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Authors and Affiliations

  1. INSERM UMR1089, University of Nantes, Centre Hospitalier Universitaire, Nantes, France

    Magalie Penaud-Budloo, Frédéric Broucque, Katell Harrouet, Mohammed Bouzelha, Sylvie Saleun, Sandy Douthe, Oumeya Adjali, Véronique Blouin & Eduard Ayuso

  2. Brammer Bio, Alachua, FL, 32615, USA

    Susan D’Costa, Sushma Ogram & Richard O. Snyder

  3. Gene Therapy Program, Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA

    Martin Lock

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  1. Magalie Penaud-Budloo
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  13. Eduard Ayuso
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Corresponding author

Correspondence to Eduard Ayuso.

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Conflict of interest

ROS is an inventor on patents related to recombinant AAV technology. ROS owns equity in a gene therapy company that is commercializing AAV for gene therapy applications. To the extent that the work in this manuscript increases the value of these commercial holdings, ROS has a conflict of interest. ML is an inventor on patents related to recombinant AAV technology. The other authors declare that they have no conflict of interest.

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Penaud-Budloo, M., Broucque, F., Harrouet, K. et al. Stability of the adeno-associated virus 8 reference standard material. Gene Ther 26, 211–215 (2019). https://doi.org/10.1038/s41434-019-0072-9

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