Sterile round bottom 96 well plates

The screening strain was grown to log phase in MHB and, when appropriate, supplemented with NaCl as noted in Results. The cells were diluted in the same growth medium to a final OD₆₀₀ of 0.01 in sterile round-bottom 96-well plates (Costar). The antibodies were added and bacteria were grown statically for 4 h at 37°C. The optical density of bacterial growth (OD₆₀₀) was measured by a plate reader after shaking the plate for 25 s. CFU was measured by serially diluting the treated bacterial culture in PBS and spotting onto agar medium.

The test strain was grown to log phase in MHB II supplemented with 0.002% Tween-80, diluted to a final OD₆₀₀ 0.01 in sterile round-bottom 96-well plates (Costar). Antibodies were added at 10 μg/mL, incubated statically at 37°C and monitored for bacterial growth after 4 hr of static incubation. A plate reader at OD₆₀₀ measured optical density of bacterial growth after shaking the plate for 25 s. Bacterial inhibition was calculated by subtracting the OD₆₀₀ value from the media control, followed by dividing that value by the no antibody control OD₆₀₀ value. Antibody activity experiments were performed as part of our high throughput antibody screening workflow and are purely descriptive.