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.

Screening and characterization of anti‐SEB peptides using a bacterial display library and microfluidic magnetic sorting

“…The two sequences have a similarity score of 25.0, which is comprised of the four consensus residues and the similarity of Thr and Ser (polar, uncharged amino acids) preceding the consensus in both sequences. The aligned consensus sequence of CHYW from this bacterial display library is very similar to the WHK and FYW consensus from peptides isolated during phage library sorting (Goldman et al, ; Soykut et al, ) and the solid phase library sequence of YYWLHH (Wang et al, ). These sequences all contain a hydrophobic Trp residue in proximity to a His residue and/or Tyr residue.…”

Screening and characterization of anti‐SEB peptides using a bacterial display library and microfluidic magnetic sorting

“…The two sequences have a similarity score of 25.0, which is comprised of the four consensus residues and the similarity of Thr and Ser (polar, uncharged amino acids) preceding the consensus in both sequences. The aligned consensus sequence of CHYW from this bacterial display library is very similar to the WHK and FYW consensus from peptides isolated during phage library sorting (Goldman et al, ; Soykut et al, ) and the solid phase library sequence of YYWLHH (Wang et al, ). These sequences all contain a hydrophobic Trp residue in proximity to a His residue and/or Tyr residue.…”

Implementation of Combinatorial Genetic and Microenvironmental Engineering to Microbial-Based Field-Deployable Microbead Biosensors for Highly Sensitive and Remote Chemical Detection

“…To this end, a peptide-presenting M13 bacteriophage capable of binding nitro compounds with high affinity was employed to increase the local concentration of nitro compounds within the microbeads, expediting the fluorescence protein expression (Figure a). The “phage display” technology has been widely used to develop a library of peptides or proteins on the bacteriophage surface, by inserting relevant genes into the phagemid, with the primary goal of identifying and studying ligand–protein interactions for drug discovery and sensor development. ,− For this study, M13 presenting WHW (Trp–His–Trp) peptides was chosen as the nitro-binding M13 (NB-M13), as previous studies have confirmed its high affinity toward both DNT and TNT (Figure S4), though the affinity toward TNT was greater than that toward DNT due to the additional nitro group interacting more favorably with the WHW motif. NB-M13 was conveniently encapsulated into the microbead by inclusion in the precursor solution prior to microbead fabrication, and the amount was controlled up to 100 μg mL –1 (further increase disrupted the gelation).…”

Evolutionary Qβ Phage Displayed Nanotag Library and Peptides for Biosensing

“…Most phage display libraries and biosensor development technologies were exclusively designed and executed with the filamentous DNA phage M13 [ 38 , 39 ]. The M13 minor coat proteins (pIII) are located only at one end of the filamentous phage and are not equally distributed upon its surface similar to in the newly developed icosahedral Qβ RNA phage [ 8 ].…”

Screening and characterization of anti‐SEB peptides using a bacterial display library and microfluidic magnetic sorting

“…The two sequences have a similarity score of 25.0, which is comprised of the four consensus residues and the similarity of Thr and Ser (polar, uncharged amino acids) preceding the consensus in both sequences. The aligned consensus sequence of CHYW from this bacterial display library is very similar to the WHK and FYW consensus from peptides isolated during phage library sorting (Goldman et al, ; Soykut et al, ) and the solid phase library sequence of YYWLHH (Wang et al, ). These sequences all contain a hydrophobic Trp residue in proximity to a His residue and/or Tyr residue.…”

Implementation of Combinatorial Genetic and Microenvironmental Engineering to Microbial-Based Field-Deployable Microbead Biosensors for Highly Sensitive and Remote Chemical Detection

“…To this end, a peptide-presenting M13 bacteriophage capable of binding nitro compounds with high affinity was employed to increase the local concentration of nitro compounds within the microbeads, expediting the fluorescence protein expression (Figure a). The “phage display” technology has been widely used to develop a library of peptides or proteins on the bacteriophage surface, by inserting relevant genes into the phagemid, with the primary goal of identifying and studying ligand–protein interactions for drug discovery and sensor development. ,− For this study, M13 presenting WHW (Trp–His–Trp) peptides was chosen as the nitro-binding M13 (NB-M13), as previous studies have confirmed its high affinity toward both DNT and TNT (Figure S4), though the affinity toward TNT was greater than that toward DNT due to the additional nitro group interacting more favorably with the WHW motif. NB-M13 was conveniently encapsulated into the microbead by inclusion in the precursor solution prior to microbead fabrication, and the amount was controlled up to 100 μg mL –1 (further increase disrupted the gelation).…”

Evolutionary Qβ Phage Displayed Nanotag Library and Peptides for Biosensing

“…Most phage display libraries and biosensor development technologies were exclusively designed and executed with the filamentous DNA phage M13 [ 38 , 39 ]. The M13 minor coat proteins (pIII) are located only at one end of the filamentous phage and are not equally distributed upon its surface similar to in the newly developed icosahedral Qβ RNA phage [ 8 ].…”

Screening and characterization of anti‐SEB peptides using a bacterial display library and microfluidic magnetic sorting

“…The two sequences have a similarity score of 25.0, which is comprised of the four consensus residues and the similarity of Thr and Ser (polar, uncharged amino acids) preceding the consensus in both sequences. The aligned consensus sequence of CHYW from this bacterial display library is very similar to the WHK and FYW consensus from peptides isolated during phage library sorting (Goldman et al, ; Soykut et al, ) and the solid phase library sequence of YYWLHH (Wang et al, ). These sequences all contain a hydrophobic Trp residue in proximity to a His residue and/or Tyr residue.…”

Implementation of Combinatorial Genetic and Microenvironmental Engineering to Microbial-Based Field-Deployable Microbead Biosensors for Highly Sensitive and Remote Chemical Detection

“…To this end, a peptide-presenting M13 bacteriophage capable of binding nitro compounds with high affinity was employed to increase the local concentration of nitro compounds within the microbeads, expediting the fluorescence protein expression (Figure a). The “phage display” technology has been widely used to develop a library of peptides or proteins on the bacteriophage surface, by inserting relevant genes into the phagemid, with the primary goal of identifying and studying ligand–protein interactions for drug discovery and sensor development. ,− For this study, M13 presenting WHW (Trp–His–Trp) peptides was chosen as the nitro-binding M13 (NB-M13), as previous studies have confirmed its high affinity toward both DNT and TNT (Figure S4), though the affinity toward TNT was greater than that toward DNT due to the additional nitro group interacting more favorably with the WHW motif. NB-M13 was conveniently encapsulated into the microbead by inclusion in the precursor solution prior to microbead fabrication, and the amount was controlled up to 100 μg mL –1 (further increase disrupted the gelation).…”

Evolutionary Qβ Phage Displayed Nanotag Library and Peptides for Biosensing

“…Most phage display libraries and biosensor development technologies were exclusively designed and executed with the filamentous DNA phage M13 [ 38 , 39 ]. The M13 minor coat proteins (pIII) are located only at one end of the filamentous phage and are not equally distributed upon its surface similar to in the newly developed icosahedral Qβ RNA phage [ 8 ].…”