title: Selection of staphylococcal enterotoxin B (SEB)-binding peptide using phage display technology.
abstract: In this study, peptides were selected to recognize staphylococcal enterotoxin B (SEB) which cause food intoxication and can be used as a biological war agent. By using commercial M13 phage library, single plaque isolation of 38 phages was done and binding affinities were investigated with phage-ELISA. The specificities of the selected phage clones showing high affinity to SEB were checked by using different protein molecules which can be found in food samples. Furthermore, the affinities of three selected phage clones were determined by using surface plasmon resonance (SPR) sensors. Sequence analysis was realized for three peptides showing high binding affinity to SEB and WWRPLTPESPPA, MNLHDYHRLFWY, and QHPQINQTLYRM amino acid sequences were obtained. The peptide sequence with highest affinity to SEB was synthesized with solid phase peptide synthesis technique and thermodynamic constants of the peptide-SEB interaction were determined by using isothermal titration calorimetry (ITC) and compared with those of antibody-SEB interaction. The binding constant of the peptide was determined as 4.2+/-0.7 x 10(5)M(-1) which indicates a strong binding close to that of antibody.
authors: Department of Food Engineering, Hacettepe University, Beytepe 06532, Ankara, Turkey.
authors: Esra Acar
authors: EA
authors: Soykut
authors: Fahriye Ceyda
authors: FC
authors: Dudak
authors: Ismail Hakki
authors: IH
authors: Boyaci
mesh_terms: D000595:Amino Acid Sequence; D017104:Bacteriophage M13 / Q000235:genetics / Q000302:isolation & purification; D002151:Calorimetry; D004768:Enterotoxins / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D020539:Sequence Analysis, Protein; D020349:Surface Plasmon Resonance
chemical_list: D004768:Enterotoxins; D019151:Peptide Library; D010455:Peptides; C031818:enterotoxin B, staphylococcal