Large-scale purification of functional AAV particles packaging the full genome using short-term ultracentrifugation with a zonal rotor.

ABSTRACT: Adeno-associated virus (AAV) vector-based gene therapy is potentially curative for various genetic diseases; however, the development of a scalable purification method for full-genome AAV vectors remains crucial to increase productivity and reduce cost of GMP production. In this study, we developed a large-scale short-term purification method for functional full-genome AAV particles by using 2-step cesium chloride (CsCl) density-gradient ultracentrifugation with a zonal rotor. The 2-step CsCl method with a zonal rotor improves separation between empty and full-genome AAV particles, reducing the ultracentrifugation time (4-5 h) and increasing the AAV volume for purification. The highly purified full-genome AAV particles were confirmed by analytical ultracentrifugation (AUC), droplet digital

SUBMITTER: Wada M 

PROVIDER: S-EPMC10457186 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature