Fig. 1

Overview of experiments assessing the influence of buffer formulation on the thermal stability and infectivity of AAV6.2FF. A In vitro assay design to evaluate the effect of buffer formulation on the infectivity and stability of AAV6.2FF-ffLuc at different temperatures and upon exposure to multiple rounds of freeze-thaw. AAV6.2FF-ffLuc vector was formulated in PBS or 0.001% PF-68-PBS and aliquoted in 1x10⁹ vg doses into 1.5 mL microfuge tubes. Samples (n=4) were subjected to incubation at -20℃, 4℃, 21℃, 37℃, and 55℃ for durations of 30 minutes (m), 1, 4, 12 or 24 hours (h) to 2, 3, or 5 days (d) or 1 or 2 weeks (w). Another set of samples (n=4) were subjected to 1 to 10 freeze-thaw cycles. All samples were evaluated for transducing activity in HEK293 cells (n=4) or for particle integrity by qPCR (n=3). B In vivo assay to investigate the effects of buffer formulation on the in vivo transducing properties of AAV6.2FF-eGFP. C57BL/6 mice (n=7) were intranasally administered 1x10¹¹ vg AAV6.2FF-eGFP with or without 0.001% PF-68. Mice were euthanized 5 weeks post vector administration and lungs were harvested, dissociated, stained for CD45, EpCAM, and 7AAD and subjected to flow cytometric analysis