Phage-displayed peptides as biosensor reagents

Abstract

This study investigated the potential to utilize phage‐displayed peptides as reagents in sensor applications. A library of random 12‐mers displayed on phage was panned against staphylococcal enterotoxin B (SEB), a causative agent of food poisoning. Nine SEB binding phage clones were isolated, all of which share the consensus sequence Trp His Lys at their amino terminus. Binding of several of these phage was shown to be inhibited when they were assayed in a competitive enzyme‐linked immunosorbent assay (ELISA) format with synthesized peptide corresponding to the peptide‐encoding region of one of the clones. Whole phage were labeled with the dye Cy5, and incorporated into fluoroimmunoassays. Labeled phage were able to detect SEB down to a concentration of 1.4 ng/well in a fluorescence‐based immunoassay. When incorporated into an automated fluorescence‐based sensing assay, Cy5‐labeled phage bound to probes coated with SEB generated a robust signal of about 10,000 pA, vs a signal of 1000 pA using a control fiber coated with streptavidin. These results demonstrate the potential for development of phage‐based sensor reagents. Copyright © 2000 John Wiley & Sons, Ltd.

The Use of Phages and Aptamers as Alternatives to Antibodies in Medical and Food Diagnostics

“…Classical ways of detection are mainly based on multiplex PCR (McLauchlin et al 2000;Mehrotra et al 2000) and immunoassays (Freed et al 1982;Wieneke 1991), which allow quantitative or qualitative measurements in less than 24 hours. A peptide displaying-phage for the optical detection of SEB with biosensor technology was developed by Goldman et al (2000). The SEB binding phage was selected from a random www.intechopen.com…”

Generation of Fab Fragment-Like Molecular Recognition Proteins against Staphylococcal Enterotoxin B by Phage Display Technology

“…Further study is needed to clarify the reason for the increased affinity for SEB. E. coli cells have been used as a host strain for the production of recombinant antibodies (7,8,29,35). The high yield of recombinant antibodies from E. coli cells is important.…”

“…Hexamer and dodecamer peptide ligands that bind to SEB have been isolated from a combinatorial peptide library (8,32,34), but the affinity of these small-molecule ligands to SEB does not seem high. However, production of a SEB-specific recombinant antibodylike protein has not been reported.…”

The Promise of Phage Display: Customized Affinity and Specificity

“…Anderson and co-workers were the first to integrate phage display with analysis (22). They selected M13 phage from a library of 12 residue peptides displayed on P3 and containing ~10 9 unique sequences for affinity to Staphylococcal enterotoxin B (SEB).…”

“…A polypeptide selected from phage display can be resynthesized or expressed with an appropriate linker and used as a receptor in a biosensor (22,26). However, retaining the phage as a display scaffold can confer important advantages.…”