An enzyme-free and label-free fluorescent biosensor for small molecules by G-quadruplex based hybridization chain reaction.

An enzyme-free and label-free fluorescent biosensor is developed by G-quadruplex-based hybridization chain reaction (HCR) for small molecules, using adenosine triphosphate (ATP) as the model. Aptamer probes for the recognition of small molecules are hybridized with blocking probes. The G-quadruplex sequences are incorporated into one of the two HCR hairpin probes. In the presence of small molecules (ATP), the formation of aptamer-ATP bioaffinity complexes induces the release of blocking probes; the released blocking probes initiate HCR and numerous G-quadruplexes along DNA nanowires are self-assembled after the HCR process. Using N-methyl mesoporphyrin IX (NMM) as the fluorophore, a "turn-on" fluorescence response can be achieved and detected as low as 15 μmol L(-1) of ATP. This biosensor is applied to detect ATP in biologic samples with satisfactory results.