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 1,000 pA using a control fiber coated with streptavidin. These results demonstrate the potential for development of phage-based sensor reagents.

Total internal reflection fluorescence (TIRF) array biosensors for biothreat agents for food safety and food defense

“…Such aptamer-beacon constructs have been used in other systems for label-free detection of proteins (Levy, Cater, & Ellington, 2005) and could potentially be transitioned into TIRF-based systems. Combinatorial peptides (peptide aptamers) are also an area of interest and have been applied to both fiber-optic evanescent wave-based systems (Goldman et al, 2000) and multiplexed array formats (Song, Stadler, Peng, & Ko Ferrigno, 2011).…”

Recent advances in recognition elements of food and environmental biosensors: A review

“…Another type of fluorescent assay is the competitive assay. This approach was used to develop a sensitive assay for the detection of Staphylococcal enterotoxin B (SEB) (Goldman et al, 2000), which is a causative agent of food poisoning and trinitrotoluene (TNT) (Goldman et al, 2003(Goldman et al, , 2002. In both these assays, fluorescent phages were used that display a peptide or scFv with specific affinity for the target.…”

Engineering protein modules for diagnostic applications

“…Peptide-based domains also offer improved stability in harsh environmental conditions such as high temperature [24]. Efforts to isolate high-affinity peptides from phage displayed libraries are now a widely used approach to screen for unstructured potent peptide binders [7, 25,26]. Recently, a new class of highly structured polypeptides has been exploited for diagnostic applications.…”