Temperature dependence of RSV inactivation.
(a) Dot blots demonstrate motavizumab recognition of antigenic site II, AM14 recognition of quaternary antigenic site V and D25 and 5C4 recognition of antigenic site Ø after RSV is incubated with a 0.025% concentration of formalin in 24-hour intervals at 25 °C (blue), 30 °C (green), 37 °C (orange) and 39 °C (red). Average antibody binding of virions is reported relative to antibody binding of a pre-F protein loading control with error bars representing standard deviation of three experiments. Significant differences between populations incubated for 0 and 72 hours are calculated by One-Way ANOVA and noted graphically (ns, not significant; *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001). Recognition of antigenic site II, present on both pre-F and post-F surfaces, is preserved after treatment with a dilute concentration of formalin at all temperatures tested. In contrast, dot blots demonstrate a temperature dependent loss of pre-F-specific mAbs AM14, D25 and 5C4 lose reactivity against virions at every temperature tested. (b) Percentage of cells infected by virus incubated with 0.025% concentration of formalin in 24-hour intervals at 4 different temperatures as in (a) by flow cytometry (Chen 2010).