摘要
Carbon nitride nanoparticles (CNNPs) have been employed as fluorescent sensing tools owing to their unique features, e.g. low cost production, high stability in water and high photoluminescence quantum yield. Here, an easy and versatile synthetic approach was exploited to design fluorescent nanoparticles with surface functionalities suitable for covalent binding to bioligands. High hydrophilic, brightly fluorescent CNNPs, rich of superficial amines, were obtained from the thermal condensation of urea and lysine (CNNP^Lys) and by tuning the precursor ratio and the heating time. Structure and size of the functionalized nanoparticles were characterized through infrared (IR) spectroscopy, transmission electron microscopy (TEM) and dynamic light scattering (DLS). Their optical properties were studied by ultraviolet-visible (UV-Vis) and fluorescence spectroscopy. The superficial primary amino groups, furnished by the lysine co-precursor, enabled for covalently linking CNNP^Lys to model proteins. The CNNP^Lys-protein conjugates excited under UV irradiation emit in the 400-450 nm visible range (quantum yield 24%). The applicability of CNNP^Lys as novel fluorescent probes was demonstrated by a fluorescence quenching assay, in which gold nanoparticles (GNPs) were attached to Staphylococcal protein A and employed to quench the CNNP^Lys fluorescence by Forster resonant energy transfer (FRET). The quenching occurred upon formation of the specific binding between the GNP-linked protein A and CNNP^Lys-tagged immunoglobulins, while the inhibition of the binding resulted in the recovery of CNNP^Lys luminescence. The synthetic strategy, based on combining a "conjugated polymer"-forming unit (urea) and a co-precursor able to provide the desired functional group (lysine), allows designing innovative materials for the development of new generation fluorescence biosensors in which easily functionalized fluorophores are needed.
