title: OneBac: platform for scalable and high-titer production of adeno-associated virus serotype 1-12 vectors for gene therapy.
abstract: Scalable and genetically stable recombinant adeno-associated virus (rAAV) production systems combined with facile adaptability for an extended repertoire of AAV serotypes are required to keep pace with the rapidly increasing clinical demand. For scalable high-titer production of the full range of rAAV serotypes 1-12, we developed OneBac, consisting of stable insect Sf9 cell lines harboring silent copies of AAV1-12 rep and cap genes induced upon infection with a single baculovirus that also carries the rAAV genome. rAAV burst sizes reach up to 5 × 10(5) benzonase-resistant, highly infectious genomic particles per cell, exceeding typical yields of current rAAV production systems. In contrast to recombinant rep/cap baculovirus strains currently employed for large-scale rAAV production, the Sf9rep/cap cell lines are genetically stable, leading to undiminished rAAV burst sizes over serial passages. Thus, OneBac combines full AAV serotype options with the capacity for stable scale-up production, the current bottleneck for the transition of AAV from gene therapy trials to routine clinical treatment.
authors.affiliation: 1 Institute of Virology , Campus Benjamin Franklin, Charité Universitätsmedizin Berlin, 12203 Berlin, Germany .
authors.forename: Mario
authors.initials: M
authors.lastname: Mietzsch
authors.forename: Sabrina
authors.initials: S
authors.lastname: Grasse
authors.forename: Catherine
authors.initials: C
authors.lastname: Zurawski
authors.forename: Stefan
authors.initials: S
authors.lastname: Weger
authors.forename: Antonette
authors.initials: A
authors.lastname: Bennett
authors.forename: Mavis
authors.initials: M
authors.lastname: Agbandje-McKenna
authors.forename: Nicholas
authors.initials: N
authors.lastname: Muzyczka
authors.forename: Sergei
authors.initials: S
authors.lastname: Zolotukhin
authors.forename: Regine
authors.initials: R
authors.lastname: Heilbronn
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D000229:Dependovirus / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors / Q000235:genetics* / Q000302:isolation & purification*; D057809:HEK293 Cells; D006367:HeLa Cells; D006378:Helper Viruses; D006801:Humans; D014161:Transduction, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D014764:Viral Proteins