Fengli Qu

In situ synthesis of palladium nanoparticle-graphene nanohybrids and their application in nonenzymatic glucose biosensors

A nonenzymatic electrochemical biosensor was developed for the detection of glucose based on an electrode modified with palladium nanoparticles (PdNPs)-functioned graphene (nafion-graphene). The palladium nanoparticle-graphene nanohybrids were synthesized using an in situ reduction process. Nafion-graphene was first assembled onto an electrode to chemically adsorb Pd(2+). And Pd(2+) was subsequently reduced by hydrazine hydrate to form PdNPs in situ. Such a PdNPs-graphene nanohybrids-based electrode shows a very high electrochemical activity for electrocatalytic oxidation of glucose in alkaline medium. The proposed biosensor can be applied to the quantification of glucose with a wide linear range covering from 10 μM to 5mM (R=0.998) with a low detection limit of 1 μM. The experiment results also showed that the sensor exhibits good reproducibility and long-term stability, as well as high selectivity with no interference from other potential competing species.

Graphene oxide quantum dots@silver core–shell nanocrystals as turn-on fluorescent nanoprobe for ultrasensitive detection of prostate specific antigen

We report a fluorescent turn-on nanoprobe for ultrasensitive detection of prostate specific antigen (PSA) based on graphene oxide quantum dots@silver (GQDs@Ag) core-shell nanocrystals. The success of this work relies on the assembly of quantities of GQDs in one GQDs@Ag probe, which makes the ratio of probe to target significantly increased and thus enables the fluorescent signal enhancement. When the silver shell was removed via oxidative etching using hydrogen peroxide (H2O2), the incorporated GQDs could be readily released and the whole process caused little change to their fluorescence performance. We tested the probe for the ultrasensitive detection of PSA based on the sandwich protocol of immunosensors. In particular, magnetic beads (MBs) were employed to immobilize anti-PSA antibody (Ab1) and acted as a separable capture probe, while GQDs@Ag was used as detection probe by linking antibody (Ab2). The developed immunosensor showed a good linear relationship between the fluorescence intensity and the concentration of PSA in the range from 1 pg/mL to 20 ng/mL with a detection limit of 0.3 pg/mL. The immunosensor used for the analysis of clinical serum samples exhibited satisfactory results, which demonstrated its potential for practical diagnostic applications. This method provides a possible solution to the application of GQDs in immunosensing and could be potentially extended to other similar systems.

Ultrasensitive electrochemical immunosensor based on horseradish peroxidase (HRP)-loaded silica-poly(acrylic acid) brushes for protein biomarker detection.

We report an ultrasensitive electrochemical immunosensor designed for the detection of protein biomarkers using horseradish peroxidase (HRP)-loaded silica-poly(acrylic acid) brushes (SiO2-SPAABs) as labels. HRP could be efficiently and stably accommodated in the three-dimensional architecture of the SiO2-SPAABs and the SiO2-SPAABs-HRP exhibited high catalytic performance towards o-phenylenediamine (OPD) oxidation in the presence of H2O2, which resulted in significant differential pulse voltammetric (DPV) response change and color change. Using human IgG (HIgG) as a model analyte, a sandwich-type immunosensor was constructed. In particular, graphene oxide (GO) and SiO2-SPAABs-HRP were used to immobilize capture antibody (Ab1) and bind a layer of detection antibody (Ab2), respectively. The current biosensor exhibited a good linear response of HIgG from 100pg/mL to 100μg/mL with a detection limit of 50pg/mL (S/N=5). The sensitivity was 6.70-fold higher than the conventional enzyme-linked immunosorbent assays. The immunosensor results were validated through the detection of HIgG in serum samples.

Novel turn-on fluorescent detection of alkaline phosphatase based on green synthesized carbon dots and MnO2 nanosheets

Using sterculia lychnophora seeds as precursors for the first time, fluorescent carbon dots (CDs) were synthesized by simple hydrothermal treatment. The quantum yield of as-synthesized CDs was 6.9% by using quinine sulfate as the reference. The fluorescence of CDs could be effectively quenched by a MnO2 nanosheet based on fluorescence resonance energy transfer (FRET). Ascorbic acid (AA) could reduce MnO2 to Mn2+ and result in the destruction of the MnO2 nanosheets, which could induce the fluorescence recovery of the CDs. In particular, alkaline phosphatase (ALP) could bio-catalyze acid 2-phosphate (AAP) hydrolysis to AA. Here, an efficient fluorescence probe based on a CDs-MnO2 nanosheet for rapid and selective detection of ALP was reported for the first time. Excellent performance for the detection of ALP was observed with high sensitivity and a detection limit of 0.4U/L owing to the low background. The detection of ALP in human serum was conducted with satisfactory results, demonstrating its potential applications in clinical diagnosis.

Hollow PDA-Au nanoparticles-enabled signal amplification for sensitive nonenzymatic colorimetric immunodetection of carbohydrate antigen 125.

A novel colorimetric immunoassay was designed for the sensitive detection of carbohydrate antigen 125 (CA125). The success of this immunoassay relies on the use of hollow polydopamine-gold nanoparticles (PDA-Au) for signal amplification to achieve sensitive nonenzymatic colorimetric detection. In particular, PDA-Au was used as a stable and sensitive label and aminated-Fe3O4 magnetic nanoparticles (Fe3O4 NPs) were employed to immobilize capture antibody (Ab1) and acted as a separable immunosensing probe. PDA-Au exhibited high catalytic performance towards p-nitrophenol reduction and thus resulted in significant color change and UV/vis signal variations. The immunoassay was performed based on sandwich protocol. As compared to pure Au nanoparticles, the signal amplification and sensitivity of PDA-Au-based assay was significantly improved. For instance, the dynamic range of the developed colorimetric immunoassay for CA125 was 0.1-100 U/mL with a detection limit of 0.1 U/mL at S/N=3. In addition, this immunoassay was also tested for the analysis of clinical serum samples, which demonstrated its potential for practical diagnostic applications.

Sensitive electrochemical immunosensor for cancer biomarker with signal enhancement based on nitrodopamine-functionalized iron oxide nanoparticles

A novel electrochemical immunosensor for sensitive detection of cancer biomarker prostate specific antigen (PSA) based on nitrodopamine (NDA) functionalized iron oxide nanoparticles (NDA-Fe(3)O(4)) is described. NDA-Fe(3)O(4) was used both for the immobilization of primary anti-PSA antibody (Ab(1)) and as secondary anti-PSA antibody (Ab(2)) label. For the preparation of the label, mediator thionine (TH) was first conjugated onto NDA-Fe(3)O(4) based on the amino groups of NDA, and then the amino group of TH was used to immobilize horseradish peroxidase (HRP) and Ab(2). Due to the high amount of NDA anchored onto Fe(3)O(4) surface, the loading of antibodies as well as mediator and enzyme onto NDA-Fe(3)O(4) was substantially increased, which increased the sensitivity of the immunosensor. The resulting immunosensor displayed a wide range of linear response (0.005-50 ng/mL), low detection limit (4 pg/mL), good reproducibility and stability. The immunosensor was used to detect the PSA contents in serum samples with satisfactory results.

Niche nanoparticle-based FRET assay for bleomycin detection via DNA scission

We describe a highly sensitive nanoparticle-based fluorescence resonance energy transfer (FRET) probe developed without using molecular fluorophores as donors and acceptors. The success of this work relies on the strategy that DNA scission was designed to occur to the probe when target presented, which enabled the fluorescence signal turn-on of graphene quantum dots (GQDs) and thus quantitative analysis. In particular, amino-modified SiO2 NPs were initially coated by GQDs to form highly emitting SiO2/GQDs, followed by conjunction with DNA functionalized gold nanoparticles (Au NPs-DNA) to form SiO2/GQDs-DNA-Au NPs composite. Owing to the FRET interactions between the GQDs and Au NPs, the fluorescence of GQDs was effectively quenched by Au NPs. When bleomycin (BLM), a model analyte, was mixed with the probe, the fluorescence signal of GQDs would be restored due to the removal of Au NPs from the SiO2/GQDs surface by DNA scission treatment with BLM in the presence of Fe (II). The current FRET probe shows a good linear relationship between the fluorescence intensity and the concentration of BLM in the range from 0.5nM to 1μM with a detection limit of 0.2nM. The probe also shows satisfactory results for the analysis of clinical serum samples. This method provides versatility to the application of GQDs in FRET biosensing and could be potentially extended to other similar systems by replacing the linker between the GQDs and Au NPs.

Pyrophosphate-regulated Zn2+-dependent DNAzyme activity: An amplified fluorescence sensing strategy for alkaline phosphatase

In this work, based on the fact that pyrophosphate (PPi) could regulate the activity of Zn(2+)-dependent DNAzyme, we for the first time report a fluorescence turn-on sensing system for alkaline phosphatase (ALP) with improved sensitivity via nonprotein-enzymatic signal amplification. A catalytic and molecular beacon (CAMB) design was employed to further improve its sensitivity. Taking advantage of the strong interactions between PPi and the Zn(2+), the cofactor Zn(2+) was caged, and the DNAzyme activity was effectively inhibited. The introduction of ALP, however, could catalyze the hydrolysis of PPi and release free Zn(2+), resulting in the activation of DNAzyme to catalyze the cleavage of the molecular beacon substrate with a remarkable increase of fluorescent signal. These optimized designs together allow a high sensitivity for ALP, with a detection limit of 20 pM observed, much lower than previously reported methods. It has also been used for detection of ALP in human serum with satisfactory results, demonstrating its potential applications in clinical diagnosis.

A universal amplified strategy for aptasensors: Enhancing sensitivity through allostery-triggered enzymatic recycling amplification

A universal amplified sensing strategy based on endonuclease was developed for designing fluorescence aptasensors. By employing hairpin-structured design for both recognition and reporter probes to decrease background signal, and a nicking endonuclease to perform target-triggered enzymatic recycling amplification, the proposed biosensor showed high sensitivity to target protein. To demonstrate the feasibility of the design, immunoglobulin E (IgE) was studied as a model target. Upon the addition of target protein, the specific formation of IgE/aptamer complex induced the releasing of the 37-mer fragment which partially hybridized with the molecular beacon (MB) probe. In the presence of endonuclease Nt.BbvCI, the MB was cleaved into two parts. Then, the released 37-mer fragment hybridized with another MB, and triggered the second cycle of cleavage, leading to an accumulation of fluorescence signals. Under the optimal conditions, a detection limit of 5 pM was obtained. The proposed sensing system was used for detection of IgE in complex biological samples with satisfactory results.

Molecular spectroscopic studies on the interaction of ferulic acid with calf thymus DNA.

The interaction between ferulic acid and calf thymus deoxyribonucleic acid (ctDNA) under physiological conditions (Tris-HCl buffer solutions, pH 7.4) was investigated by UV-Vis spectroscopy, fluorescence spectroscopy, DNA melting techniques, and viscosity measurements. Results indicated that a complex of ferulic acid with ctDNA was formed with a binding constant of K(290K)=7.60×10(4) L mol(-1) and K(310K)=4.90×10(4) L mol(-1). The thermodynamic parameters enthalpy change (ΔH°), entropy change (ΔS°) and Gibbs free energy (ΔG°) were calculated to be -1.69×10(4) J mol(-1), 35.36 J K(-1) mol(-1) and -2.79×10(4) J mol(-1) at 310 K, respectively. The acting forces between ferulic acid and DNA mainly included hydrophobic interaction and hydrogen bonds. Acridine orange displacement studies revealed that ferulic acid can substitute for AO probe in the AO-DNA complex which was indicative of intercalation binding. Thermal denaturation study suggested that the interaction of ferulic acid with DNA could result in the increase of the denaturation temperature, which indicated that the stabilization of the DNA helix was increased in the presence of ferulic acid. Spectroscopic techniques together with melting techniques and viscosity determination provided evidences of intercalation mode of binding for the interaction between ferulic acid and ctDNA.