Purification of adenovirus and adeno-associated virus: comparison of novel membrane-based technology to conventional techniques.
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.
Duffy
A M
AM
Department of Medicine and Regenerative Medicine Institute (REMEDI), National University of Ireland, Galway, Ireland.
O'Doherty
A M
AM
O'Brien
T
T
Strappe
P M
PM
D000256:Adenoviridae; D001709:Biotechnology; D002460:Cell Line; D000229:Dependovirus; D016166:Free Radical Scavengers; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000302:isolation & purification*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D013194:Staining and Labeling; D013482:Superoxide Dismutase / Q000032:analysis / Q000235:genetics; D019076:Transgenes; D014773:Virology / Q000379:methods
D016166:Free Radical Scavengers; D011993:Recombinant Fusion Proteins; D013482:Superoxide Dismutase