Significant Enhancement of Adeno‐Associated Virus Vector Yield for Gene Therapy Using Multidimensional Optimization Strategies

Gene therapy holds great promise for the treatment of currently incurable diseases, enabled by engineered viral vectors that facilitate precise in vivo and ex vivo gene delivery. Since its discovery in 1965, adeno-associated virus (AAV) has attracted substantial attention in virology owing to its unique intrinsic properties. The engineering of the first recombinant AAV (rAAV) in 1984 marked a pivotal transition, catalyzing its exploration as a therapeutic vector for genetic disorders. However, current challenges facing rAAV-based gene therapy, particularly inefficient vector production and prohibitively high manufacturing costs, necessitate systematic optimization. This review delineates multidimensional enhancement strategies for scalable rAAV manufacturing through systematic investigation across three pivotal dimensions: (i) genetic modification of AAV and its auxiliary components, (ii) modulation of the relationship between viral replication and the host cell cycle, and (iii) optimization of upstream production and downstream purification processes. These integrated approaches collectively establish a robust theoretical framework for developing standardized protocols to produce high-titer, clinical-grade rAAV virion preparations.