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In this work, the potential application of the fluorescence dye Thioflavin-T (ThT), which can specifically bind to amyloid, as a powerful tool for monitoring secondary structural transitions of silk fibroin (SF) induced by pH in low solution concentrations was examined. Results showed that ThT emission intensities substantially increased when pH decreased from 6.8 to 4.8. This increase may be ascribed to conformational transitions from random coil to β-sheet. The morphology and secondary structure of SF were also investigated via TEM, AFM and circular dichroism spectroscopy. The information obtained herein can be utilized not only for the development of convenient and efficient noninvasive method for monitoring the assembly behavior of SF in aqueous solution but also for in vitro fluorescence imaging. [ABSTRACT FROM AUTHOR]

In this research, we designed a label-free fluorometric turn-on assay for trypsin and inhibitor screening, based on a spherical cationic gemini surfactant ethylene-bis (dodecyl dimethyl ammonium bromide) (EDAB)/heparin/Nile red (NR) supramolecular assembly system. The introduction of gemini surfactant EDAB as template greatly enhanced its salt resistance and resulted in the supramolecular assemblies with diameters ranging from 20 to 100 nm. The fluorometric assay for trypsin was performed by firstly disassembling with protamine (a heparin-binding protein) and then re-assembling through hydrolysis of protamine. The disassembly and reassembly of the system resulted in a turn-off first and then a turn-on behavior of the corresponding fluorescence. The overall processes were characterized by fluorescence spectra, TEM measurements and zeta potential tests. The detection level of this assembly system for trypsin was as low as 4.2 ng mL−1. Also, the EDAB/heparin/NR assembly could be used to screen the trypsin inhibitors. The assembly system was easily-fabricated and cost-effective, but also exhibited good salt tolerance in NaCl solution at the concentration of 0–500 mM. At last, the supramolecular assembly was successfully applied to detect trypsin in human urine, demonstrating its great potential on clinical diagnosis applications. [ABSTRACT FROM AUTHOR]

Highly soluble fluorescent pyrene derivative with substantially improved fluorescence intensity in aqueous buffer was obtained viaPEGylation strategy. The highly soluble PEGylated pyrene (PEO-Py) non-covalently adsorbed onto the surface of gold nanoparticles (Au NPs) to form dyads with quenched fluorescence due to highly efficient energy transfer between PEO-Py and Au NPs. The PEO-Py/Au NPs dyads were used for the sensitive turn-on fluorescent detection of biothiols. The fluorescence of PEO-Py was restored by the addition of cysteine (Cys), indicating that Cys can modulate the energy transfer between PEO-Py and Au NPs. This phenomenon then allowed for the sensitive detection of Cys with a limit of detection (LOD) of 11.4 nM. The linear range of determination of Cys was from 1.25 × 10−8to 2.25 × 10−7M. None of the other amino acids found in proteins showed obvious interference with the determination. It was important to note that the detection sensitivity of the PEO-Py/Au NPs system was more than 5-fold improved compared with the Py/Au NPs system. In addition, other biothiol molecules, such as glutathione, could also be detected by this sensor system. The method was also successfully applied to the determination of the total content of aminothiols in human plasma. Therefore an easily prepared, inexpensive, high solubility fluorescent probe has been realized and is also expected to detect other biological analytes of interest. [ABSTRACT FROM AUTHOR]

Highly soluble fluorescent pyrene derivative with substantially improved fluorescence intensity in aqueous buffer was obtained viaPEGylation strategy. The highly soluble PEGylated pyrene (PEO-Py) non-covalently adsorbed onto the surface of gold nanoparticles (Au NPs) to form dyads with quenched fluorescence due to highly efficient energy transfer between PEO-Py and Au NPs. The PEO-Py/Au NPs dyads were used for the sensitive turn-on fluorescent detection of biothiols. The fluorescence of PEO-Py was restored by the addition of cysteine (Cys), indicating that Cys can modulate the energy transfer between PEO-Py and Au NPs. This phenomenon then allowed for the sensitive detection of Cys with a limit of detection (LOD) of 11.4 nM. The linear range of determination of Cys was from 1.25 × 10−8to 2.25 × 10−7M. None of the other amino acids found in proteins showed obvious interference with the determination. It was important to note that the detection sensitivity of the PEO-Py/Au NPs system was more than 5-fold improved compared with the Py/Au NPs system. In addition, other biothiol molecules, such as glutathione, could also be detected by this sensor system. The method was also successfully applied to the determination of the total content of aminothiols in human plasma. Therefore an easily prepared, inexpensive, high solubility fluorescent probe has been realized and is also expected to detect other biological analytes of interest. [ABSTRACT FROM AUTHOR]