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Purification of adenovirus and adeno-associated virus: comparison of novel membrane-based technology to conventional techniques

Gene Therapy volume 12, pages S62–S72 (2005)Cite this article

Abstract

Adenovirus (Ad) and Adeno-associated virus (AAV) are efficient gene delivery systems; manipulation of the wild-type genome allows their use as vectors for the overexpression of desirable transgenes. Generation and purification of such viral vectors can be labour intensive, costly and require specialized equipment, but a new generation of membrane-mediated ion exchange kits for purification of recombinant virus may facilitate this process. Here, we examine the yields, transgene expression and purity of preparations of Ad and AAV purified using commercially available kits in comparison to other established techniques for purification of recombinant viral vectors. We demonstrate comparable results for Ad and AAV respectively in all parameters investigated, with a substantial reduction in purification time for the kit-based technology. Such approaches are attractive methods for small-scale purification of recombinant Ad and AAV viral vectors.

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Acknowledgements

We thank Science Foundation Ireland (SFI), Health Research Board Ireland and Higher Education Authority for their financial support and also Martina Harte (NUIG) and Vecteurotrain, for their guidance in establishing the adenoviral purification protocol. We thank Dr T Athanasopoulos, Dr J Harris and Dr G Dickson for guidance and advice with iodixanol gradient and heparin column purification of AAV, Dr D Grimm and Dr J Kleinschmidt for plasmid pDG and Dr N Madigan for cloning of PON1. We also thank Dr D Draganov for anti-hPON1 antibody.

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  1. Department of Medicine and Regenerative Medicine Institute (REMEDI), National University of Ireland, Galway, Ireland

    A M Duffy, A M O'Doherty, T O'Brien & P M Strappe

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  1. A M Duffy
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  2. A M O'Doherty
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  3. T O'Brien
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  4. P M Strappe
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Duffy, A., O'Doherty, A., O'Brien, T. et al. Purification of adenovirus and adeno-associated virus: comparison of novel membrane-based technology to conventional techniques. Gene Ther 12 (Suppl 1), S62–S72 (2005). https://doi.org/10.1038/sj.gt.3302616

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