As a popular controllable-released carrier, intelligent hydrogels are often used in drug delivery and disease therapeutics. Meanwhile, benefit from the mimic-enzyme activity performance, Fe-N-C nanozymes have been widely used in sensing and analysis. However, the combination of intelligent hydrogels with specific degradability and Fe-N-C nanozymes with enhanced activity in one system to achieve controllable and sensitive detection is rare. Herein, we combine intelligent hydrogel with mimic peroxidase activity enhanced Fe-N-C nanozymes to construct a ratiometric fluorescence probe for sensitive detection of hyaluronidase (HAase). The modification of copper ions has been proved to enhance the mimic enzyme activity of Fe-N-C nanozymes greatly. Cu2+ modified Fe-N-C nanozymes were embedded in hyaluronic acid hydrogel. In the presence of HAase, the HA hydrogel structure was hydrolyzed and released Cu2+-Fe-N-C nanozymes gradually. The released Cu2+-Fe-N-C nanozymes are used to catalyze the hydrogen peroxide system so that o-phenylenediamine is oxidized to orange fluorescent 2, 3-diaminophenolazine (DAP). Due to the electrostatic interaction, the fluorescence resonance energy transfer can occur between the negatively charged copper nanoclusters emitted by 430 nm and the positively charged DAP emitted by 560 nm. The activity of HAase was monitored according to the ratio of fluorescence intensity at 560 nm and 430 nm (F560/F430). The linear range of this method is 0-10.0 U/ml and the detection limit is 0.43 U/mL (S/N = 3). This strategy has been further applied to biological samples successfully.
Keywords: Intelligent hydrogels; Metal nanoclusters; Nanozymes; Ratiometric fluorescence.
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