Figure 4.

Testing of AAV6 produced at 3L scale using either Bac-RepCap or the SGMO Helper

(A) SDS-PAGE and Coomassie blue stained gel of purified AAV6 from 3L production run. The individual Cap (VP1, VP2, and VP3) proteins are indicated on the panel. The proteolytic cleavage fragment observed with AAV6 produced using Bac-RepCap is indicated by an asterisk. AAV8 produced using the 293 production system (293 Reference) was also included for comparison. The capsid ratio for each production run is reported in Table 1. (B) In vitro potency of AAV6 produced using 293 cells or at 3L scale in Sf9 cells (Bac-RepCap and SGMO Helper) on HepG2 cells. HepG2 cells were transduced using the indicated MOI and, after 5 days, cell supernatants were assayed for α-Gal A activity. The nmol/h/mL α-Gal A activity was calculated based on fluorescent activity using a standard curve. Each virus sample was assessed for potency in triplicate, with each of the replicates being assayed in duplicate, and the standard deviation of the six data points was used to plot error bars.