Monoclonal antibodies disrupt specific activities of Coa. (A and B) Association of Coa with human prothrombin or fibrinogen was measured by ELISA and perturbed with increasing concentrations of affinity-purified 5D5, affinity-purified 3B3, polyclonal antibodies (α-Coa), or IgG1 isotype control. (C) Calcium-chelated mouse blood was inoculated with S. aureus Newman (1 × 106 CFU) in the presence of 3 µM of 5D5, 3B3, polyclonal antibodies (α-Coa), or IgG1 isotype control. Samples were incubated at room temperature and monitored for coagulation. (D) Rabbit EDTA-plasma was mixed with SYTO9 stained S. aureus Newman (1 × 107 CFU) in the presence of 3 µM of 5D5, 3B3, polyclonal antibodies (α-Coa), or IgG1 isotype control. Samples were incubated at room temperature for 10 min, analyzed by fluorescence microscopy, and quantified by calculating means ± SEM from 12 fields of microscopic view. Statistical significance was assessed with one-way ANOVA and Bonferroni post-test. *, P < 0.01; **, P < 0.001; ***, P < 0.0001.
