Stability of the adeno-associated virus 8 reference standard material

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. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution ACCESS OPTIONS Access

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subscriptions * Read our FAQs * Contact customer support REFERENCES * Aucoin MG, Perrier M, Kamen AA. Critical assessment of current adeno-associated viral vector production and

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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,

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

32615, USA Susan D’Costa, Sushma Ogram & Richard O. Snyder * Gene Therapy Program, Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA, 19104,

USA Martin Lock Authors * Magalie Penaud-Budloo View author publications You can also search for this author inPubMed Google Scholar * Frédéric Broucque View author publications You can also

search for this author inPubMed Google Scholar * Katell Harrouet View author publications You can also search for this author inPubMed Google Scholar * Mohammed Bouzelha View author

publications You can also search for this author inPubMed Google Scholar * Sylvie Saleun View author publications You can also search for this author inPubMed Google Scholar * Sandy Douthe

View author publications You can also search for this author inPubMed Google Scholar * Susan D’Costa View author publications You can also search for this author inPubMed Google Scholar *

Sushma Ogram View author publications You can also search for this author inPubMed Google Scholar * Oumeya Adjali View author publications You can also search for this author inPubMed Google

Scholar * Véronique Blouin View author publications You can also search for this author inPubMed Google Scholar * Martin Lock View author publications You can also search for this author

inPubMed Google Scholar * Richard O. Snyder View author publications You can also search for this author inPubMed Google Scholar * Eduard Ayuso View author publications You can also search

for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Eduard Ayuso. ETHICS DECLARATIONS 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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Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE 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 Download citation * Received: 18 December 2018 * Revised: 05 March 2019 * Accepted: 08 March 2019 * Published: 29

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