Enzyme‐Instructed Self‐Assembly Enabled Monomer–Excimer Transition to Construct Higher Ordered Luminescent Supramolecular Assembly for Activity‐based Bioimaging

Abstract It is challenging to construct high‐performing excimer‐based luminescent analytic tools at low molecular concentrations. We report that enzyme‐instructed self‐assembly (EISA) enables the monomer–excimer transition of a coumarin dye ( Cou ) at low molecular concentrations, and the resulting higher ordered luminescent supramolecular assemblies (i.e., nanofibers) efficiently record the spatiotemporal details of alkaline phosphatase (ALP) activity in vitro and in vivo. Cou was conjugated to short self‐assembly peptides with a hydrophilic ALP‐responsive group. By ALP triggering, EISA actuated a nanoparticles–nanofibers transition at low peptide concentrations followed by monomer–excimer transition of Cou . Analysis of structure–property relationships revealed that the self‐assembly motif was a prerequisite for peptides to induce the monomer–excimer transition of Cou . Luminescent supramolecular nanofibers of pYD ( LSN‐ pYD ) illuminated the intercellular bridge of cancer cells and distinguished cancer cells (tissues) from normal cells (tissues) efficiently and rapidly, promising potential use for the early diagnosis of cancer. This work extends the functions of EISA and provides a new application of supramolecular chemistry.