Solid-Phase Ligand-Binding Assay

This assay was used to confirm the ability of GapA to bind to selected F. tularensis proteins and was conducted as described previously (Pavkova et al., 2017). Briefly, the purified recombinant proteins were coated onto a 96-well high binding microtiter plate overnight and then incubated with recombinant GapA (0.25–2.5 μg/ml). The amount of GapA bound to the proteins was determined spectrophotometrically (450 nm) in an enzyme-linked immunosorbent assay using anti-GapA antibody (Apronex, Vestec, Czechia).