Similarly, the diluted culture was dispensed to Cell Culture Treated EasYFlasks, 175 cm2 (Nunc, Roskilde, Denmark) at two-time points (12 h, 24 h) in triplicates and incubated at 37 °C in a static condition. After incubation, the used media along with the planktonic cells were washed off and the flask was rinsed twice with sterile PBS to remove the settled planktonic cells. The biofilm cells were then scraped off, pooled in sterile ice-cold PBS, pelleted and stored at -80 °C till protein isolation.