title: Serotype-independent method of recombinant adeno-associated virus (AAV) vector production and purification.
abstract: A variety of gene transfer strategies have been developed to treat inherited, degenerative, and acquired diseases. Among the different vector systems developed so far, recombinant adeno-associated viral (AAV) vectors have shown notable benefits, including prolonged gene expression, transduction of both dividing and nondividing cells, and a lack of pathogenicity caused by wild-type infections. Thanks to these features, the use of AAV vectors as a gene transfer tool has increased dramatically during the past several years, and several recent clinical trials have used AAV vectors. However, AAV vectors are more complicated to produce than are other viral vectors. With steady advances toward clinical application, much effort has been made to isolate novel AAV serotypes and to develop methods for their efficient, scalable, and versatile production and purification. Here we review state of the art methods for AAV vector production and purification, which we have refined in our laboratory.
authors.affiliation: Department of Biochemistry and Molecular Biology, Division of Gene Therapy Research Center for Advanced Medical Technology, Nippon Medical School, Tokyo, Japan. kmiyake@nms.ac.jp
authors.forename: Koichi
authors.initials: K
authors.lastname: Miyake
authors.forename: Noriko
authors.initials: N
authors.lastname: Miyake
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Yamazaki
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimada
authors.forename: Yukihiko
authors.initials: Y
authors.lastname: Hirai
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000145:classification / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012703:Serotyping; D014161:Transduction, Genetic / Q000379:methods*
chemical_list: D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins