An efficient and scalable process for helper-dependent adenoviral vector production using polyethylenimine-adenofection.

摘要: Safety requirements for adenoviral gene ther- apy protocols have led to the development of third generation vectors commonly called helper-dependent (HDVs). HDVs demonstrated a high therapeutic potential; however, poor efficiency and reliability actual production process hampers further large-scale clinical evaluation this new vector. The current HDV methods involve preliminary rescue step through transfection adherent cell cultures by an plasmid followed helper adenovirus (HV) infection. Amplification serial co-infection comple- mentary cells allows increase in titer. Using HEK293 FLP/frt system suspension culture, alter- native protocol transfection/infection proce- dure was evaluated. In work, adenofection uses linked HV with help polyethy- lenimine (PEI) has shown outperform standard producing higher yield. influence complex composition on examined statistical design. optimized amplification conditions were successively performed generate at 3 L bioreactor scale. Following only two passages, up 1.44 � 10 8 infectious units/mL culture generated, which corresponded 26% total particles produced. This strat- egy, realized reduced duration therefore probability vector recombina- tion introducing cost-effective protocol, ensuring high-quality stock. Biotechnol. Bioeng. 2009;102: 800-810. 2008 Wiley Periodicals, Inc.