Polymer‐Templated Perylene‐Probe Noncovalent Self‐Assembly: A New Strategy for Label‐Free Ultrasensitive Fluorescence Turn‐On Biosensing

A new label-free fluorescence turn-on strategy for highly sensitive biosensing has been developed. A negatively charged perylene probe was synthesized. Polycations could induce aggregation of the perylene probe through noncovalent interactions and the fluorescence of the probe's monomer was efficiently quenched. Upon addition of a single-stranded nucleic acid, competitive binding of the negatively charged nucleic acid (a polyanion) to the cationic polymer resulted in the release of a monomer and thus a turn-on fluorescence signal was detected. Without the use of any amplification techniques, a detection limit of 2 pM DNA was obtained. Based on these results, an assay strategy for the highly sensitive detection of alkaline phosphatase (ALP) activity has been demonstrated. λ Exonuclease (λ exo) could degrade 5'-phosphorylated single-stranded DNA. However, when the DNA sample was treated with ALP, the phosphate functional group was removed by ALP and it could no longer be degraded by λ exo. Binding of the DNA to the perylene probe-polycation complex resulted in a turn-on fluorescence signal, which could be used for sensing of ALP. The method is highly sensitive, a limit of detection as low as 0.02 mU mL(-1) ALP was obtained. Our method is simple, convenient, highly sensitive, and inexpensive.