Figure 5.

Filtration assay on Amicon centrifugal filters

AAV8-GFP were produced in HEK293 (A and B) or in Sf9 cells (C and D) and purified by the CsCl gradient method. AAV vectors were loaded on a centrifugal filter of 100 kDa MWCO that does not allow the AAV particles to pass through. A synthetic miR-19 was spiked before filtration in the Sf9-derived AAV sample, as control of free miRNA. Vector genomes (A and C) were quantified by qPCR before concentration, and in the concentrated (C) and flowthrough (FT) fractions. The quantity of residual miRNAs (B and D) was quantified by RT-qPCR in the C and FT fractions after centrifugation.