Stimuli-responsive azobenzene-quantum dots for multi-sensing of dithionite, hypochlorite, and azoreductase.

A new fluorescence turn-on sensing platform has been developed applicable for sensitive profiling of multiple chemical and biological analytes, using azobenzene-quantum dot as a new stimuli-responsive optical nanoprobe. An azobenzene-carrying compound bis [4, 4'-(dithiophenyl azo)-1, 3-benzenediamine] (DTPABDA) is for the first time reported to be used for conjugation with CdSe/ZnS core/shell quantum dots (QDs) via the ligand exchange reaction. Due to the photo-induced electron-transfer (PET) effect, the electron-withdrawing azobenzene groups of DTPABDA can significantly cause the photoluminescence (PL) of QDs quenched. The QDs' PL can be subsequently reignited by the removal of azo moiety cleavable through three types of specific reactions: the dithionite reduction, hypochlorite oxidation, and azoreductase enzymatic catalysis, respectively. By monitoring of reaction-induced recovery of FL signals at 560 nm with an excitation of 450 nm, such azobenzene-QDs conjugates served as a new nanoprobe enabling the fluorescence turn-on sensing of dithionite, hypochlorite, and azoreductase with high sensitivity, broad linear range, and good selectivity. The successful detection of target analytes in real samples reveals the potential of our method in practical applications, such as biosensing, environmental and industrial monitoring. Graphical abstract A new stimuli-responsive fluorescence probe is reported for the sensitive detection of sodium dithionite, hypochlorite, and azoreductase. The probe consists of QDs with an azobenzene-carrying compound as a ligand. The fluorescence of QDs could be quenched by the azo group and subsequently recovered via the removal of azo group by these three compounds, resulting in the "turn-on" sensing of these compounds with high sensitivity, broad linear range, and good selectivity. The successful detection of azoreductase in serum samples reveals the practical use of this method.

Fluorescent Probe Encapsulated in SNAP-Tag Protein Cavity To Eliminate Nonspecific Fluorescence and Increase Detection Sensitivity.

Despite the promising improvements made recently on fluorescence probes for the detection of enzymes and reactive small molecules, two fundamental problems remain: weaker fluorescence of many dyes in aqueous buffers and strong nonspecific signals in samples containing high protein levels. In this paper, we introduce a novel fluorescent probe encapsulated in protein cavity (FPEPC) concept as demonstrated by SNAP-tag protein and three environment-sensitive fluorescence probes to overcome these two problems. The probes were constructed by following the current probe design for enzymes and reactive small molecules but with an additional benzylguanine moiety for selective SNAP-tag conjugation. The SNAP-tag conjugated probes achieved quantitative nitroreductase and hydrogen sulfide detection in blood plasma, whereas analyte concentrations were overestimated up to 700-fold when bare fluorescent probes were employed for detection. Furthermore, detection sensitivity was increased dramatically, as our probes displayed 390-fold fluorescence enhancement upon SNAP-tag conjugation, in stark contrast to the weak fluorescence of the free probes in aqueous solutions. Compared with the conventional approaches where fluorescent probes are encapsulated into polymers and nanoparticles, our simple and general approach successfully overcame many key issues such as dye leakage, long preparation steps, inconsistent dye-host ratios, difficulty in constructing in situ in a complex medium, and limited application to detect only small metabolites.

Fluorescent Probe Encapsulated in Avidin Protein to Eliminate Nonspecific Fluorescence and Increase Detection Sensitivity in Blood Serum.

Quantitative detection of trace amounts of a biomarker in protein rich human blood plasma using fluorescent probes is a great challenge as the real signal is usually obscured by nonspecific fluorescence. This problem occurs because most of the fluorescent dyes bind very tightly with blood proteins to produce a large fluorescence increase, resulting in overestimation of the biomarker concentrations and false positive diagnosis. In this paper, we report that biotinylated fluorescent probes encapsulated in avidin protein can generate very specific fluorescence in blood serum by blocking out nonspecific dye-protein interactions. We applied our novel probe design to detect two different types of biomolecules, hydrogen sulfide and nitroreductase. Our Avidin conjugated probes achieved quantitative analyte detection in blood serum; whereas concentrations were overestimated up to 320-fold when bare fluorescent probes were employed. As compared to conventional approaches where fluorescent probes are encapsulated into polymers and nanoparticles, our simple approach successfully overcomes many key issues such as dye leakage, long preparation steps, inconsistent dye-host ratios, difficulty in constructing in situ in a complex medium, and limited application to detect only small metabolites.