We report on the synthesis and characterization of highly fluorescent gold nanoclusters (Au-NCs) from a gold precursor using a single-step chemical reduction in the presence of human adult hemoglobin (Hb). The conformational changes of Hb before and after cluster formation were studied by various spectroscopic techniques. The Au-NCs in Hb display blue fluorescence with a peak centered at 450 nm (photoexcited at 365 nm) and a quantum yield of 2.8 %. The Au-NCs exhibit excellent photostability and long-term stability, and can be applied in the pH range 5-12 even in the presence of high electrolyte concentrations. The Au-NCs in Hb can act as a highly sensitive and selective fluorescent turnoff probe for Cu(II) ion. The observed reversible fluorescence recovery of Hb-AuNCs/Cu(II) aggregates was exploited to develop a selective and sensitive turn-on fluorescence assay for His. Under optimized conditions, the probe gives a fluorescent response that is linear in the 0.1 to 20.0 μM concentration range of Cu(II), with a limit of detection of 28 nM. The probe for His, in turn, has a linear range in the 1-21 μM concentration range, and the limit of detection is 0.6 μM.
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