Kiichi Hirohata, Yuki Yamaguchi, Takahiro Maruno, Risa Shibuya, Tetsuo Torisu, Takayuki Onishi, Hideto Chono, Junichi Mineno, Yuan Yuzhe, Kiyoko Higashiyama, Kyoko Masumi-Koizumi, Kazuhisa Uchida, Takenori Yamamoto, Eriko Uchida, Takashi Okada, Susumu Uchiyama
Adeno-associated virus (AAV) vectors are produced as a mixture of the desired particle (full particle, FP), which is filled with the designed DNA, product-related impurities such as particle without DNA (empty particle, EP), and aggregates. Cesium chloride or iodixanol equilibrium density gradient ultracentrifugation (DGE-UC) has been used for the purification of AAV vectors. DGE-UC can separate FP from impurities based on the difference in their buoyant densities. Here, we report the applications and limitations of equilibrium density gradient analytical ultracentrifugation (DGE-AUC) using a modern AUC instrument that employs DGE-UC principles for the characterization and quantitation of AAV vectors...
January 2, 2024: Analytical Chemistry