A novel ratiometric fluorescent tyrosinase assay is developed based on hybrid nano-assembly of gold nanocluster and tyrosine-containing peptides. The AuNCs@YCY nano-probe (AYNP) is fabricated through the hydrophobic interactions and π-π stacking between the tyrosine residues of the Tyr-Cys-Tyr tripeptide (YCY) and the ligands on the surfaces of AuNCs under the near-isoelectric pH value. The resulted AYNP shows distinct fluorescence responses, spontaneous turn-on of the blue emission and turn-off of the near-infrared emission, with a single wavelength excitation. It is demonstrated that the enhancement and quenching are due to the production of pheomelanin and dopaquinone structures, respectively, induced by tyrosinase oxidation. The internal referencing system provides the tyrosinase assay with superior sensitivity and a detection limit as low as 6.3 U L-1 could be achieved. The experimental results also demonstrate the excellent selectivity, good photo-stability, and both in vitro and cellular applications of AYNP. This assay technique is low-cost, easy to prepare, and shows excellent potential as a novel melanoma clinical diagnostic platform and a tyrosinase inhibitor screening tool.
Keywords: Cell imaging; Melanoma; Ratiometric fluorescent probe; Tyrosinase assay.
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