Yuan-Di Zhao

The interface behavior of hemoglobin at carbon nanotube and the detection for H2O2

Direct electrochemistry of hemoglobin (Hb) is observed at carbon nanotube (CNT) interface. The adsorbing Hb can transfer electron directly at CNT interface compared with common carbon material. The heterogeneous electron transfer rate constant k of Hb can be calculated as 0.062s(-1), the transfer coefficient alpha is 0.21 and the average surface coverage of Hb on CNT surface is 3.58 x 10(-9) +/- 2.7 x 10(-10)mol/cm(2). It is found that the adsorbing Hb still keeps its catalytic activity to H(2)O(2). This sensor was used to detect H(2)O(2). The apparent Michaelis-Menten constant is calculated as 6.75 x 10(-4)molL(-1).

Anodic oxidation of hydrazine at carbon nanotube powder microelectrode and its detection

Carbon nanotube powder microelectrodes (CNTPMEs) were used to study the anodic oxidation of hydrazine at Carbon nanotube (CNT)-the novel carbon material. It was found that the electrochemical behaviours were greatly improved at CNTs, indicating that the anodic oxidation could be catalyzed at CNTs. The kinetics parameters of this process were calculated, the heterogeneous electron transfer rate constant k was 0.0019 cm s(-1), (1-alpha)n(alpha) was 0.22. The CNTPMEs were also found with high sensitivity for hydrazine detection, could be used as hydrazine sensors.

Distance-Dependent Metal-Enhanced Quantum Dots Fluorescence Analysis in Solution by Capillary Electrophoresis and Its Application to DNA Detection

Here the distance dependence of metal-enhanced quantum dots (QDs) fluorescence in solution is studied systematically by capillary electrophoresis (CE). Complementary DNA oligonucleotides-modified CdSe/ZnS QDs and gold nanoparticles (Au NPs) were connected together in solution by the hybridization of complementary oligonucleotides, and a model system (QD-Au) for the study of metal-enhanced QDs fluorescence was constructed, in which the distance between the QDs and Au NPs was controlled by adjusting the base number of the oligonucleotide. In our CE experiments, the metal-enhanced fluorescence of the QDs solution was only observed when the distance between the QDs and Au NPs ranged from 6.8 to 18.7 nm, and the maximum enhancement by a factor of 2.3 was achieved at 11.9 nm. Furthermore, a minimum of 19.6 pg of target DNA was identified in CE based on its specific competition with the QD-DNA in the QD-Au system. This work provides an important reference for future study of metal-enhanced QDs fluorescence in solution and exhibits potential capability in nucleic acid hybridization analysis and high-sensitivity DNA detection.

High-sensitivity quantum dot-based fluorescence resonance energy transfer bioanalysis by capillary electrophoresis

Here a new method for high-sensitivity quantum dot (QD)-based fluorescence resonance energy transfer (FRET) bioanalysis was developed. In this method, capillary electrophoresis (CE) with fluorescence detection was applied. The FRET system consisted of water-soluble 532-nm emitting CdTe QDs donor and 632-nm emitting CdSe/ZnS QDs acceptor which were covalently conjugated with mouse IgG and goat anti-mouse IgG, respectively. The bio-affinity between antigen and antibody brought two kinds of QDs close enough to make the FRET happen between them. In the CE experiments, highly efficient separation of donor-acceptor immunocomplexes was obtained, and the process of FRET was monitored. Results showed that FRET efficiency obtained by CE (38.56-69.58%) improved substantially in comparison with that obtained by ensemble measurement (12.77-52.37%). The high efficient separation of donor-acceptor immunocomplexes and the possible conformation change of antigen and antibody, contributes to the lower analysis uncertainty (variance) and higher FRET efficiency obtained in CE and consequentially, this makes the analysis of FRET more sensitive. This novel CE-based technique can be easily extended to other FRET system based on QDs and may have potential application in the study of biomolecule conformation change.

Simultaneous detection of dual single-base mutations by capillary electrophoresis using quantum dot-molecular beacon probe

Here a novel capillary electrophoresis (CE) for simultaneous detection of dual single-base mutations using quantum dot-molecular beacon (QD-MB) probe is described. Two QD-MB probes were designed using 585 and 650-nm emitting CdTe QDs which were covalently conjugated to MBs with different DNA oligonucleotide sequences by amide linkage and streptavidin-biotin binding, respectively. The hybridizations of QD-MB probes with different DNA targets were then monitored by CE, and results indicated that the two QD-MB probes specifically hybridized with their complementary DNA sequences, respectively. Target DNA identification was observed to have a high sensitivity of 16.2 pg in CE. Furthermore, the simultaneous detection of dual single-base mutations in a given DNA oligonucleotide was successfully achieved in CE using above two QD-MB probes. This novel CE-assisted QD-MB biosensor offers a promising approach for simultaneous detection of multiple single-base mutations, and exhibits potential capability in the single nucleotide polymorphism (SNP) analysis and high-sensitivity DNA detection.

Influence of quantum dot's quantum yield to chemiluminescent resonance energy transfer

The resonance energy transfer between chemiluminescence donor (luminol-H2O2 system) and quantum dots (QDs, emission at 593 nm) acceptors (CRET) was investigated. The resonance energy transfer efficiencies were compared while the oil soluble QDs, water soluble QDs (modified with thioglycolate) and QD-HRP conjugates were used as acceptor. The fluorescence of QD can be observed in the three cases, indicating that the CRET occurs while QD acceptor in different status was used. The highest CRET efficiency (10.7%) was obtained in the case of oil soluble QDs, and the lowest CRET efficiency (2.7%) was observed in the QD-HRP conjugates case. This result is coincident with the quantum yields of the acceptors (18.3% and 0.4%). The same result was observed in another similar set of experiment, in which the amphiphilic polymer modified QDs (emission at 675 nm) were used. It suggests that the quantum yield of the QD in different status is the crucial factor to the CRET efficiency. Furthermore, the multiplexed CRET between luminol donor and three different sizes QD acceptors was observed simultaneously. This work will offer useful support for improving the CRET studies based on quantum dots.

Electrocatalytic oxidation of phytohormone salicylic acid at copper nanoparticles-modified gold electrode and its detection in oilseed rape infected with fungal pathogen Sclerotinia sclerotiorum.

Salicylic acid (SA) is a biological substance that acts as a phytohormone and plays an important role in signal transduction in plants. It is important to accurately and sensitively detect SA levels. A gold electrode modified with copper nanoparticles was used to assay the electrocatalytic oxidation of salicylic acid. It was found that the electrochemical behavior of salicylic acid was greatly improved at copper nanoparticles, indicating that anodic oxidation could be catalyzed at copper nanoparticles. And the pH had remarkable effect on the electrochemical process, a very well-defined oxidation peak appeared at pH 13.3 (0.2M NaOH). The kinetics parameters of this process were calculated and the heterogeneous electron transfer rate constant (k) was determined to be 1.34x10(-3)cms(-1), and (1-alpha)n(alpha) was 1.22. The gold electrode modified with copper nanoparticles could detect SA at a higher sensitivity than common electrodes. The electrode was used to detect the SA levels in oilseed rape infected with the fungal pathogen Sclerotinia sclerotiorum. The results showed that the SA concentration reached a maximum during the 10th-25th hours after infection. This result was very similar to that determined by HPLC, indicating that the gold electrodes modified with copper nanoparticles could be used as salicylic acid sensors.

Targeted quantum dots fluorescence probes functionalized with aptamer and peptide for transferrin receptor on tumor cells

Quantum dots (QDs) fluorescent probes based on oligonucleotide aptamers and peptides with specific molecular recognition have attracted much attention. In this paper, CdSe/ZnS QDs probes for targeted delivery to mouse and human cells using aptamer GS24 and peptide T7 specific to mouse/human transferrin receptors were developed. Capillary electrophoresis analyses indicated that the optimal molar ratios of QDs to aptamer or peptide were 1:5. Fluorescence and confocal microscope imaging revealed QD-GS24 and QD-T7 probes were able to specifically recognize B16 cells and HeLa cells respectively. Quantitative flow cytometry analysis indicated the transportation of QD-GS24 or QD-T7 into cells could be promoted by corresponding free transferrin. Transmission electron microscopy confirmed the uptake of probes in cells and the effective intracellular delivery. MTT assay suggested the cytotoxicity of probes was related to the surface ligand, and aptamer GS24 (or peptide T7) could reduce the cytotoxicity of probes to a certain degree. The study has great significance for preparing QDs fluorescent probes using non-antibody target molecules.

The oxidation and reduction behavior of nitrite at carbon nanotube powder microelectrodes

Carbon nanotube electrodes were fabricated using powder microelectrode method, and the carbon nanotube powder microelectrodes (CNTPMEs) were characterized by the electro-oxidation and electro-reduction of nitrite. It was found that the kinetics of oxidation and reduction were greatly improved at CNTs compared with that at conventional graphite, indicating that CNTs could catalyze the electrochemical process of nitrite. The kinetic parameters of these process at CNTs were calculated, i.e. k was 0.593 cm s−1, and (1-α)nα was 0.501±0.018 for the nitrite oxidation. This CNTPME was also used as a nitrite carbon nanotube sensor, and the results showed that the detection limit was 8 μM.

Electrocatalytic activity of salicylic acid on the platinum nanoparticles modified electrode by electrochemical deposition

Platinum nanoparticles (PNP) were deposited on the Pt disk electrode by electrochemical deposition, it was found that salicylic acid (SA) had good electrocatalytic oxidation response at the prepared Pt nanoparticles modified Pt disk electrode (PNP/Pt). Then the modified electrode was used to detect SA in weak alkali condition, it was found that the response current was 9.2 times that of bare Pt disk electrode, and the linear range of detection was 2.0x10(-5) to 5.0x10(-4)M. After that, the electrode was adopted to detect the SA in Zuguangsan, one kind of Chinese medicine, the results showed that the PNP/Pt electrode has low detection limit, high repeatability and stability as SA sensor, could be applied in the detection of SA reliably.