Enzyme-instructed self-assembly enabled fluorescence light-up for alkaline phosphatase detection

Alkaline phosphatase (ALP) exists in both normal and pathological tissues. Spatiotemporal variations in ALP levels can reveal its potential physiological functions and changes that occur during pathological conditions. However, it is still challenging to exploit fluorescent probes that can measure ALP activity under good spatial and temporal resolutions. Herein, enzyme-instructed self-assembly (EISA) was used to construct a high-performing analytical tool (MN-pY) to probe ALP activity. MN-pY alone (free state) showed negligible fluorescence but presented an almost 13-fold increase in fluorescence intensity in the presence of ALP (assembly state). Mechanism study indicated the increase in fluorescence intensity was due to hydrogelation and formation of supramolecular fibrils, mainly consisting of dephosphorylated MN-Y. The dephosphorylation and further fibrillation of MN-pY could induce the formation of a “hydrophobic pocket”, leading to a further increase in fluorescence intensity. Moreover, MN-pY could selectively illuminate HeLa cells with a higher ALP expression but not LO2 cells with lower ALP levels, promising a potential application in cancer diagnosis. [Display omitted] •Enzyme-instructed peptide assembly (EISA) was employed to construct high-performing fluorescence tool (MN-pY).•MN-pY presented great performance responding to ALP both in vitro and at the cellular level.•Good selectivity to report ALP activity was observed.•Microenvironment variation around fluorophore due to peptide assmebly contributes to fluorescence amplification.