Analytical Ultracentrifugation as an Approach to Characterize Recombinant Adeno-Associated Viral Vectors

AUC distinguishes partially filled from empty and full capsids.

“AUC has two advantages for the quantification of the content ratio: it is highly repeatable, at only 2% CV, and distinguishes partially filled from empty and full capsids.”

AUC is highly repeatable at 2% CV.

“AUC has two advantages for the quantification of the content ratio: it is highly repeatable, at only 2% CV, and distinguishes partially filled from empty and full capsids.”

AUC is limited to purified samples without residual impurities or excipients and requires prior titration into the linear range 2 × 10^12-5 × 10^12 cp/mL.

“Sensitivity of the detectors and the risk of induced density gradients limit AUC to purified samples without residual impurities or excipients, and prior titration into the small linear range of the detector, 2 × 1012–5 × 1012 cp/mL, is needed.”

In a single experiment, AUC assesses presence of aggregates and quantifies relative content of empty, partially filled, and full capsids.

“In a single experiment, AUC assesses the presence of aggregates and quantifies the relative content of empty, partially filled, and full capsids.”

AUC requires large sample volumes (400-500 μL) at high concentrations around 2 × 10^12-5 × 10^12 vg/mL, representing significant loss of unrecoverable material during early production stages.

“In general, AUC requires large sample volumes, 400–500 μL, at high concentrations around 2 × 1012–5 × 1012 vg/mL, representing a significant loss of unrecoverable material during early production stages.”