A hexamer peptide ligand that binds selectively to staphylococcal enterotoxin B: isolation from a solid phase combinatorial library

Abstract

By screening a solid-phase combinatorial peptide library, a short peptide ligand, YYWLHH, has been discovered that binds with high affinity and selectivity to staphylococcal enterotoxin B (SEB), but only weakly to other SEs that share sequence and structural homology with SEB. Using column affinity chromatography with an immobilized YYWLHH stationary phase, it was possible to separate SEB quantitatively from Staphylococcus aureus fermentation broth, a complex mixture of proteins, carbohydrates and other biomolecules. The immobilized peptide was also used to purify native SEB from a mixture containing denatured and hydrolyzed SEB, and showed little cross-reactivity with other SEs. To our knowledge this is the first report of a highly specific short peptide ligand for SEB. Such a ligand is a potential candidate to replace antibodies for detection, removal and purification strategies for SEB.

Depletion and fractionation technologies in plasma proteomic analysis

“…Combinatorially generated peptides specific for plasma proteins have been used to selectively capture plasma ligands [31]. Peptides from these libraries can have affinities in the nanomolar range with high specificity [32,33]. Commercially, ProMetic Life Sciences has successfully utilized Mimetic™ ligands to deplete abundant plasma proteins at a large scale [115].…”

Thermodynamic and structural analysis of interactions between peptide ligands and SEB

“…Similarly another SEB binding (www.interscience.wiley.com) DOI:10.1002DOI:10. /jmr.1003 Research Article peptide was screened and selected by Wang et al (2004) using a solid phase combinatorial library. The binding specificity and affinity of this peptide was investigated by an HPLC flow analysis and affinity purification of SEB molecule was successfully realized in a column that SEB binding peptides used as affinity agents.…”

Affinity Chromatography: Principles and Applications

“…one-bead-one-peptide libraries) (Lam et al, 1991). Since then, peptide sequences have been isolated for a wide range of targets (Casey et al, 2008) and have been used to purify staphylococcal enterotoxin B (Wang et al, 2004), -tryptase (Schaschke et al, 2005), and -cobratoxin (Byeon & Weisblum, 2004). The main advantages of using peptides as affinity ligands are their low cost and stability.…”

“…Peptide affinity ligands are typically identified using one of two techniques (Wang et al, 2004); biological combinatorial peptide libraries (e.g phage-displayed libraries) (Cwirla et al, 1990;Devlin et al, 1990;Smith & Scott, 1993) or solid-phase combinatorial libraries (e.g. one-bead-one-peptide libraries) (Lam et al, 1991).…”