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Adeno-Associated Viruses

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Viral Vectors for Gene Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 737))

Abstract

Adeno-associated virus (AAV) vectors have evolved over the past decade as a particularly useful gene ­vector for in vivo applications. In contrast to oncoretro- and lentiviral vectors, this vector stays essentially episomal after gene transfer, making it safer because of the absence of insertional mutagenesis. AAV’s non-pathogenicity is a further advantage. For decades, this vector could only be produced at a small scale for research purposes and, eventually, used at very small doses for clinical studies, because only transfection methods were available, which have limited scalability. However, since the development of scalable production methods, this bottleneck is resolved and, from a technical point of view, large quantities of AAV vectors can be produced, opening the possibility of using AAV vectors for whole body treatments in gene therapy trials. This chapter presents the basic principles of small- and large-scale production procedures as well as detailed procedure of small-scale production, purification, and analytical protocols for AAV vectors. In Chapter 10, the reader will find a large-scale production method based on the use of the insect cell/baculovirus system.

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Acknowledgements

These protocols are based on the protocols used at the 3rd Eurolabcourse: Advanced industrial methods for production, purification and characterisataion of gene vectors (see http://www.easco.org/adminarea/upload/files/0306_0720_EVRY2004Report.pdf) (organizers: O.-W. Merten, M. Mezzina and G. Waksmann), Evry/F, 14–26 June 2004.

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  1. Généthon, Evry, France

    Otto-Wilhelm Merten

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  1. Mauro Mezzina
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  2. Otto-Wilhelm Merten
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Correspondence to Otto-Wilhelm Merten .

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  1. Généthon, rue de l'Internationale 1, Évry CX 2, 91002, France

    Otto-Wilhelm Merten

  2. Chemical & Bioprocess Engineering, University College Dublin, Belfield, Dublin, 4, Ireland

    Mohamed Al-Rubeai

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Mezzina, M., Merten, OW. (2011). Adeno-Associated Viruses. In: Merten, OW., Al-Rubeai, M. (eds) Viral Vectors for Gene Therapy. Methods in Molecular Biology, vol 737. Humana Press. https://doi.org/10.1007/978-1-61779-095-9_9

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