Fig. 4. Optimized thin film matrix enhances adenovirus stability at elevated temperatures and under environmental stressor conditions.

Films containing 1.25 × 10¹² vp were prepared in batch and either stored in controlled environmental chambers held at 37°C (A), 4°C (B), and 20°C (C) for 84 days or subjected to a series of 16 freeze-thaw (F/T) cycles (D). Replicates (at least three per time point) were reconstituted, and live virus concentration was assessed by a standard infectious titer assay (19). Virus was placed at the same concentration in two standard liquid formulations, and infectious titers under each storage condition were also assessed for comparison. In each panel, control A formulation consisted of 100 mM PBS (pH 7.4) and 10% glycerol, and control B formulation consisted of 100 mM PBS alone. In each panel, data represent the average ± SD. *P < 0.05, ***P < 0.001, two-tailed Student’s t test. Formulations are summarized in the figure according to the numbers listed in table S1.