High-Throughput, High-Sensitivity Peptide Functional Assay Utilizing Colorimetric Polydiacetylene

Membrane-active peptides, including antimicrobial peptides, cell-penetrating peptides, and ionophores, often require time-consuming and costly techniques for functional characterization, creating a bottleneck in drug screening. In this study, we present a high-throughput, high-sensitivity peptide functional assay utilizing the mechanochromic polymer polydiacetylene (PDA). We demonstrate that optimizing the composition of PDA can significantly enhance its assay performance, increasing the reaction speed by 4-fold and the sensitivity by 2.6-fold. Moreover, the optimized PDA assay effectively captured the cooperative effect, offering superior performance compared to that of conventional assays.