Figure 6.

ELEBA evaluation of the abilities of BSA (A,E for control), brevinin-2SSb (B,F), ranatuerin-2SSa (C,G), and granuliberin-SSa (D,H) to bind to the bacterial endotoxins LPS (A–D) and LTA (E–H). Serially diluted brevinin-2SSb, ranatuerin-2SSa, or granuliberin-SSa were coated on 96-well microplate wells and incubated with biotinylated LPS (biotin-LPS) or biotinylated LTA (biotin-LTA) for 4 h at room temperature. Peptide-bound biotin-LPS or biotin-LTA was detected by incubation with HRP-labeled streptavidin followed by incubation with the ELISA POD substrate in the TMB solution, and A₄₅₀ of the reaction products was measured. Columns and vertical bars represent means and SEM, respectively (n = 4). In panels (B,C,F,H), values with the same letters are not significantly different (p ≥ 0.05). A schematic diagram for the ELEBA is presented in Figure S3. BSA, bovine serum albumin; LPS, lipopolysaccharide; LTA, lipoteichoic acid; ELEBA, enzyme-linked endotoxin binding assay; HRP, horseradish peroxidase; ELISA, enzyme-linked immunosorbent assay; POD, peroxidase; TMB, 3,3’,5,5’-tetramethyl-benzidine.