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Methods Mol Biol. 2019;1937:91-99. doi: 10.1007/978-1-4939-9065-8_5.

AAV Production Using Baculovirus Expression Vector System.

Author information

1
University of Versailles Saint-Quentin en Yvelines, Montigny-le-Bretonneux, France.
2
SQY Therapeutics SARL, Noisy-le-Roi, France.
3
University of Versailles Saint-Quentin en Yvelines, Montigny-le-Bretonneux, France. rachid.benchaouir@uvsq.fr.
4
Centre Scientifique de Monaco, Monaco, Monaco. rachid.benchaouir@uvsq.fr.
5
SQY Therapeutics SARL, Noisy-le-Roi, France. rachid.benchaouir@uvsq.fr.

Abstract

Gene transfer and gene therapy are powerful approaches for many biological research applications and promising avenues for the treatment of many genetic or cancer diseases. The most efficient gene transfer tools are currently derived from viruses. Among them, the recombinant adeno-associated viruses (AAVs) are vectors of choice for many fundamental and therapeutic applications. The increasing number of clinical trials involving AAVs demonstrates the need to implement production and purification processes to meet the quantitative and qualitative demands of regulatory agencies for the use of these vectors in clinical trials. In this context, the rise of production levels on an industrial scale appeared essential. The introduction, in 2002, of an AAV process using a baculovirus expression vector system (BEVS) has circumvented this technological lock. The advantage of BEVS in expanding the AAV production in insect cells has been to switch the process to bioreactor systems, which are the ideal equipment for scaling up. We describe here a method for producing AAV vectors using the BEVS which can be easily used by research laboratories wishing to overcome the difficulties associated with the scaling up of production levels. The method provides sufficient quantities of AAV vectors to initiate preclinical projects in large animal models or for research projects where a single batch of vectors will consolidate the repeatability and reproducibility of in vitro and especially in vivo experimental approaches.

KEYWORDS:

AAV vectors; Baculovirus; Downstream process; Production; Purification; Upstream process

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