Comparative analysis of the burst size and cell transduction efficiency of the rAAV5 vectors produced in 293 or Sf9 cells. (A) The yields of rAAV5 preparations were quantified as benzonase-resistant genomic particles (gp) per cell by qPCR using rAAV genome-specific primers. rAAV burst sizes of three experiments are displayed as mean±standard deviation. Please note the logarithmic scale of the y axis. (B) Analysis of the transduction efficiencies of rAAV5 vectors derived from freeze–thaw supernatants as determined in HeLa C12 cells (MOI: 10,000). Transduction efficiencies of rAAV5s prepared in 293 were arbitrarily set to 1.0, while those of rAAV5 vectors derived from Sf9 cells are displayed as percentage thereof. The experiments were performed in triplicates and are displayed as mean±standard deviation. (C) Silver staining gel analysis of AVB-chromatography-purified rAAV5 preparations separated on 8% SDS-PAGE with 5×109 gp loaded per lane. (D) Analysis of rAAV5 transduction as described in (B), except that highly purified rAAV5 vector preparations were used (MOI: 1000). Data on clone 7 (ΔRBE) are added for comparison, and details are discussed in the context of Fig. 3. RBE, Rep binding element.