High-purity AAV vector production utilizing recombination-dependent minicircle formation and genetic coupling.

ABSTRACT: Triple transfection of HEK293 cells is the most widely used method for producing recombinant adeno-associated virus (rAAV), a leading gene delivery vector for human gene therapy. Despite its tremendous success, this approach generates several vector-related impurities that could potentially compromise the safety and potency of rAAV. In this study, we introduce a method for high-purity AAV vector production utilizing recombination-dependent minicircle formation and genetic coupling (AAVPure^Mfg). Compared with traditional triple transfection, AAVPure^Mfg substantially improves vector purity by reducing prokaryotic DNA contaminants by 10- to 50-fold and increasing the full capsid ratio up to threefold. Mechanistically, Bxb1-mediated excision of the transgene cassette gene

SUBMITTER: Liu H 

PROVIDER: S-EPMC12162853 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature