TESSA Platform Delivers High-Quality, High-Titre, Multi Serotype rAAV Stocks

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A significant challenge for Adeno-associated virus (AAV) -mediated gene transfer has been the capacity to produce recombinant AAV vectors of high quality, at high-titre and at large scale. OXGENE has addressed these challenges by developing a proprietary Tetracycline-Enabled Self-Silencing Adenovirus (TESSA) system that simultaneously provides Adenovirus (Ad) helper functions and encoded AAV Cap/Rep. When cells carrying an AAV ITR-flanked transducing genome are infected with TESSA-CapRep high-titre rAAV stocks are produced that are essentially free of contaminating TESSA vector.

We have now examined the ability of TESSA to support production of a diverse range of AAV serotypes. The Cap coding region of AAV serotypes 1-9 were independently engineered into the E1 region of TESSA and the quality, physical and infectious titre of the rAAV produced by this system were characterised. For all rAAV serotypes, physical titres and infectious titres were invariably greater than that observed for conventional plasmid-based rAAV systems. For these serotypes, the ratio of full to empty particles is greatly improved. Moreover, for some AAV serotypes, the TESSA system delivered infectious titres 1000-fold greater than the plasmid-based helper-free systems.

Importantly, TESSA-derived rAAV serotypes were competent for reporter gene and therapeutic gene transfer into a multiplicity of human cell types in vitro and a variety of model animal tissues in vivo. OXGENE have therefore developed a versatile system that can be applied to a range of naturally occurring AAV serotypes, and also to experimentally engineered AAV capsids with novel tissue targeting phenotypes. This system provides unique opportunities for robust high-quality scale up of rAAV manufacture that will reduce costs and facilitate rapid progression to clinical application of novel rAAV-based gene therapies.
Lead Scientist, Viral Design & Discovery
OXGENE, a WuXi Advanced Therapies Company