Msf recombinant proteins of the three predominant Msf SVs associated with hyperinvasive isolates exhibit differential binding to vitronectin.
ELISA Immulon 2HB plates were coated overnight with 3 μM of either Msf recombinant proteins (rMsf_SV-149-189, blue; rMsf_SV-249-186, green; or rMsf_SV-549-189, red) or purified Opc (yellow) protein, prior to blocking with 3% BSA. The plates were subsequently overlaid with 0.2 μg vitronectin and vitronectin binding was detected using anti-Vn monoclonal antibody 8E6 and anti-mouse alkaline phosphatase IgG (secondary antibody). SIGMAFAST™ p-Nitrophenyl phosphate tablets were used to detect alkaline phosphatase activity in the ELISA and the absorbance was measured at 405 nm. Results are the average of three independent experiments; p-values (≤ 0.01) indicated with * or **. Error bars show standard error between replicates.