The Challenges of Recombinant Adeno-associated Virus Manufacturing: Alternative Use of Adeno-associated Virus Plasmid/Liposome Complexes for Gene Therapy Applications

Recombinant adeno-associated viruses (rAAVs) have strong potential as safe and efficient transducing vectors for a variety of gene therapy applications. Wild-type AAV can infect a wide range of mammalian cell types, and, in the absence of adenovirus or herpes virus, integrates its genome into that of the host (Berns and Bohensky 1987). One major obstacle to the widespread use of rAAV is the lack of simple methods to efficiently manufacture purified recombinant virus at high titer (Rolling and Samulski 1995). Several improvements have been described (Flotte et al. 1995; Lebkowski et al. 1988; Mamounas et al. 1995), however, current methods still involve multiple, variable, time consuming procedures which result in low yields of AV and are not suitable for scale up for widespread application.