Dual genetically encoded phage-displayed ligands

Abstract

M13 bacteriophage display presents polypeptides as fusions to phage coat proteins. Such phage-displayed ligands offer useful reagents for biosensors. Here, we report a modified phage propagation protocol for the consistent and robust display of two different genetically encoded ligands on the major coat protein, P8. The results demonstrate that the phage surface reaches a saturation point for maximum peptide display.

Keywords: Bacterial transformation; ELISA; Phage; Phage display.

Copyright © 2014 Elsevier Inc. All rights reserved.

Figures

Figure 1

Agarose (1%) gel electrophoresis confirms the presence of both phagemids from dual display phage samples.

Figure 2

Phage-based ELISAs demonstrating the binding affinity of dual-displayed phage. (A) Phage generated with similar ligand sequences targeting PSMA. The inset compares a 1:1 mixture of phage-1 and -2, to phage-12 and -21. (B) Phage generated with dissimilar ligand sequences targeting PSMA. (C) Phage-based sandwich ELISA demonstrating simultaneous binding to both BSA and PSMA. Stop-4, helper phage packaging the phagemid DNA, serves as the negative control. Error bars indicate standard error (n=3).