Hemolytic Activity

The hemolysis assay was carried out by inoculating mammalian erythrocytes obtained from fresh defibrinated sheep and mouse blood (Thermo Fisher Scientific, Waltham, MA, United States) with the peptides. Briefly, peptides were incubated at 37°C with a 4% erythrocyte suspension with a twofold serial dilution series of peptides ranging from 512 to 1 μg/ml for 2 h. PBS and 1% nonionic detergent Triton X-100^TM (Sigma-Aldrich, St. Louis, MO, United States) diluted in PBS served as negative and positive controls, respectively. Cytolysis was evaluated by the measurement of absorption using a microplate reader at 550 nm. Experiments were done in triplicate. The hemolysis is derived from the following calculation:

A represents the absorbance of samples treated with peptide, while A_PBS and A_Triton indicate the absorbance of the negative and positive controls, respectively.