Peptide self-assembly assisted signal labeling for an electrochemical assay of protease activity

Peptide self-assembly holds tremendous promise for a range of applications in chemistry and biology. In the work reported here, we explored the potential functions of peptide self-assembly in electrochemical bioanalysis by developing a peptide self-assembly assisted signal labeling strategy for assaying protease activity. The fundamental principle of this assay is that target-protease-catalyzed specific proteolytic cleavage blocks self-assembly between the probe peptide and signal peptide, thus preventing the signal labeling of electroactive silver nanoparticles on the electrode surface, which in turn causes the electrochemical signal to decrease. Using trypsin as an example protease target, the linear range of this assay was found to be 1 ng mL^-1 to 100 mg mL^-1, and its detection limit was 0.032 ng mL^-1, which are better than the corresponding parameters for previously reported assays. Further experiments also highlighted the good selectivity of the assay method and demonstrated its usability when applied to serum samples. Therefore, this report not only introduces a valuable tool for assaying protease activity, but it also promotes the utilization of peptide self-assembly in electrochemical bioanalysis, as this approach has great potential for practical use in the future.