Macrocyclic Phage Display for Identification of Selective Protease Substrates.

ABSTRACT: Traditional methods for identifying selective protease substrates have primarily relied on synthetic libraries of linear peptides, which offer limited sequence and structural diversity. Here, we present an approach that leverages phage display technology to screen large libraries of chemically modified cyclic peptides, enabling the identification of highly selective substrates for a protease of interest. Our method uses a reactive chemical linker to cyclize peptides on the phage surface, while simultaneously incorporating an affinity tag and a fluorescent reporter. The affinity tag enables capture of the phage library and subsequent release of phages expressing optimal substrates upon incubation with a protease of interest. The addition of a turn-on fluorescent reporter allows direct quant

SUBMITTER: Faucher FF 

PROVIDER: S-EPMC12314892 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature