Currently, there is no rapid and unbiased screening method to identify new classes of chemical probes from highly diverse pools of candidate molecules. We have recently developed a phage display methodology that allows to directly screen for specific substrate probes for a given protease target, by chemically inserting a selectivity-directing P1 site. Our approach makes use of a reactive linker which allows to form a cyclic peptide on the phage surface while simultaneously introducing a defined a P1 cleavage site and an affinity tag to immobilize the phage on beads. Using this approach, we identified cyclic peptides that are cleaved by FAP, a therapeutically relevant serine hydrolase. Our strategy enables a rapid, unbiased screening to identify new classes of highly selective substrates for diverse protease targets.

We are expanding this technology to other protein classes and different types of chemical probes.

References

  • F Faucher, S Lovell, M Cosco, M Bertolini, M Bogyo* & M Barniol-Xicota*. Macrocyclic Phage Display Approach for Direct Selective Protease Substrate Identification. GRC Chemistry and Biology of Peptides. Oxnard (USA). [Poster]
  • Marta Barniol-Xicota. Selective Substrate Identification Using Chemically Modified Phage Display. Short talk presented at the German Peptide Symposium. Jena (Germany). August 2023
  • S Lovell, M Bogyo* & M Barniol-Xicota*. GRC Chemistry and Biology of Peptides. Oxnard (USA).