Molecular machine and gold/graphene quantum dot hybrid based dual amplification strategy for voltammetric detection of VEGF165.

Graphene quantum dots (GQDs) were prepared via pyrolysis of citric acid and glutamic acid, then reacted with chlorauric acid to form a gold/graphene quantum dot hybrid (Au/GQD), and finally connected with hairpin DNA probe 1 (H1) and thionine (Thi). The H1-Au/GQD-Thi composite is found to be a viable redox probe for electrochemical and aptamer-based determination of vascular endothelial growth factor VEGF165. A dual amplification strategy is employed based on the use of molecular machine and the Au/GQD. Each single VEGF165 molecule can bind two DNA probes via specific aptamer-target recognition to produce a molecular machine. Surface-tethered hairpin DNA 2 (H2) hybridizes with the molecular machine through proximity effect, and the prelocked toehold domain of H2 becomes exposed. This part binds to H1-Au/GQD-Thi to release the molecular machine which then moves to the neighboring H2 upon which a surface programmatic chain reaction is initiated. By continuous molecular machine travel, many H1-Au/GQD-Thi probes are present on the gold electrode surface. This implies an efficient signal amplification capability. The Au/GQD based redox probes in-situ catalyzes the redox activity of thionine and further enhances the detection signal. The aptasensor exhibits ultrahigh sensitivity and selectivity for VEGF165. The square wave voltammetric signal, best measured at -0.18 V vs. Ag/AgCl, increases linearly in the 1.0 fM to 120 pM VEGF165 concentration range, and the detection limit is 0.3 fM. Conceivably, the method may be applied to other target proteins if the corresponding high-affinity aptamers are available. Graphical abstract This study report one dual amplification strategy for ultrasensitive electrochemical detection of VEGF165 based on gold-graphene quantum dot hybrid (Au/GQD) and bipedal molecular machine (BMM) powered surface programmatic chain reaction (SPCR).

Pentaethylenehexamine and d-penicillamine co-functionalized graphene quantum dots for fluorescent detection of mercury(II) and glutathione and bioimaging.

Pentaethylenehexamine and d-penicillamine co-functionalized graphene quantum dots (PEHA-GQD-DPA) was made via one two-step thermal pyrolysis. The resulting PEHA-GQD-DPA is composed of the graphene sheets with an average size of 3.16 nm and the rich of functional groups. It gives an ultra strong fluorescence emission with the fluorescent quantum yield of 90.91% and sensitive and selective optical response towards Hg2+. The fluorescence intensity linearly decreases with the increase of Hg2+ in the range of 1.0 × 10-10-2 × 10-4 M with the detection limit of 4.6 × 10-11 M (S/N = 3). No species tested interfere with detection of Hg2+. The fluorescence quenched by Hg2+ can be well recovered by glutathione. The fluorescence intensity linearly increases with the increase of glutathione in the range of 5 × 10-8-2.5 × 10-6 M with the detection limit of 1.7 × 10-8 M (S/N = 3). The PEHA-GQD-DPA as a fluorescence probe has been successfully applied in determination of Hg2+ in natural water and glutathione in human serum and SW480 cell imaging.

Improved activity and thermo-stability of the horse radish peroxidase with graphene quantum dots and its application in fluorometric detection of hydrogen peroxide.

Graphene quantum dots (GQDs) have received extensive concern in many fields such as optical probe, bioimaging and biosensor. However, few reports refer on the influence of GQDs on enzyme performance. The paper reports two kinds of graphene quantum dots (termed as GO-GQDs and N,S-GQDs) that were prepared by cutting of graphene oxide and pyrolysis of citric acid and l-cysteine, and their use for the horse radish peroxidase (HRP) modification. The study reveals that GO-GQDs and N,S-GQDs exhibit an opposite effect on the HRP performance. Only HRP modified with GO-GQDs offers an enhanced activity (more than 1.9 times of pristine enzyme) and thermo-stability. This is because GO-GQDs offer a larger conjugate rigid plane and fewer hydrophilic groups compared to N,S-GQDs. The characteristics can make GO-GQDs induce a proper conformational change in the HRP for the catalytic performance, improving the enzyme activity and thermo-stability. The HRP modified with green luminescent GO-GQDs was also employed as a biocatalyst for sensing of H2O2 by a fluorometric sensor. The colorless tetramethylbenzidine (TMB) is oxidized into blue oxidized TMB in the presence of H2O2 by the assistance of HRP/GO-GQDs, leading to an obvious fluorescence quenching. The fluorescence intensity linearly decreases with the increase of H2O2 concentration in the range from 2×10-9 to 2×10-4M with the detection limit of 6.8×10-10M. The analytical method provides the advantage of sensitivity, stability and accuracy compared with present H2O2 sensors based on the pristine HRP. It has been successfully applied in the determination of H2O2 in real water samples. The study also opens a new avenue for modification of enzyme activity and stability that offers great promise in applications such as biological catalysis, biosensing and enzyme engineering.

Synthesis of nitrogen-doped activated graphene aerogel/gold nanoparticles and its application for electrochemical detection of hydroquinone and o-dihydroxybenzene.

Graphene aerogel materials have attracted increasing attention owing to their large specific surface area, high conductivity and electronic interactions. Here, we report for the first time a novel strategy for the synthesis of nitrogen-doped activated graphene aerogel/gold nanoparticles (N-doped AGA/GNs). First, the mixture of graphite oxide, 2,4,6-trihydroxybenzaldehyde, urea and potassium hydroxide was dispersed in water and subsequently heated to form a graphene oxide hydrogel. Then, the hydrogel was dried by freeze-drying and reduced by thermal annealing in an Ar/H2 environment in sequence. Finally, GNs were adsorbed on the surface of the N-doped AGA. The resulting N-doped AGA/GNs offers excellent electronic conductivity (2.8 × 10(3) S m(-1)), specific surface area (1258 m(2) g(-1)), well-defined 3D hierarchical porous structure and apparent heterogeneous electron transfer rate constant (40.78 ± 0.15 cm s(-1)), which are notably better than that of previous graphene aerogel materials. Moreover, the N-doped AGA/GNs was used as a new sensing material for the electrochemical detection of hydroquinone (HQ) and o-dihydroxybenzene (DHB). Owing to the greatly enhanced electron transfer and mass transport, the sensor displays ultrasensitive electrochemical response to HQ and DHB. Its differential pulse voltammetric peak current linearly increases with the increase of HQ and DHB in the range of 5.0 × 10(-8) to 1.8 × 10(-4) M for HQ and 1 × 10(-8) to 2.0 × 10(-4) M for DHB. The detection limit is 1.5 × 10(-8) M for HQ and 3.3 × 10(-9) M for DHB (S/N = 3). This method provides the advantage of sensitivity, repeatability and stability compared with other HQ and DHB sensors. The sensor has been successfully applied to detection of HQ and DHB in real water samples with the spiked recovery in the range of 96.8-103.2%. The study also provides a promising approach for the fabrication of various graphene aerogel materials with improved electrochemical performances, which can be potentially applied in biosensors, electrocatalysis, and energy storage/conversion devices.