*2.5. Minimal Inhibitory Concentration (MIC)*

The MIC was conducted using the liquid broth inhibition assay as previously described [36]. MHB was used as a medium for bacterial growth, and bacteria (either MRSA SAP0017, *P. aeruginosa* PA14, *E. coli* ATTC489 or *A. baumannii* 5015) were suspended at ∼<sup>1</sup> × 106 CFU mL<sup>−</sup>1. The peptide conjugates were incubated with the bacterial cultures at various concentrations for ~18 h prior determination of bacterial growth optical density at 600 nm (OD600). The MIC was defined as the lowest concentration that led to no growth in the wells. All values are modal values of three independent experiments.

#### *2.6. Biofilm Biomass and Biofilm Eradication Assays*

The ability of the designed conjugates to decrease the biomass of bacterial biofilms was assessed in 96-well microtitre plates using the crystal violet staining assay as described previously [37]. A bacterial suspension of 1 × <sup>10</sup><sup>7</sup> CFU mL−<sup>1</sup> was used to establish biofilms prior to treatment with the peptides. Appropriate nutrient media for optimal biofilm formation were used, including TSB supplemented with 1% glucose for MRSA SAP0017, BM2 glucose for *P. aeruginosa* PA14, and LB media supplemented with 1% glucose for *E. coli* ATTC489 and *A. baumannii* 5015. In the crystal violet-based assay, untreated bacterial culture and sterile media were used as positive and negative controls, where the % of biofilm biomass was considered 100 and 0%, respectively. The % of biomass in the treated samples was determined relative to the positive and negative controls. A biofilm eradication assay [17] was also performed, for which the residual viable biofilm cell count (CFU mL<sup>−</sup>1) was determined. Experiments were conducted in triplicate, in three independent experiments, and average values were presented with ± standard deviations.
