Guobin Mao

Aptamer-functionalized CdTe:Zn2+ quantum dots for the detection of tomato systemin

With the advantages of excellent optical properties and biocompatibility, DNA-functionalized quantum dots (QDs) have been widely applied in biosensing and bioimaging. Systemin is an important class of plant peptide hormone that was first identified in plants. In this paper, we have synthesized aptamer-functionalized Zn2+ doped CdTe QDs through a facile one-pot hydrothermal route, and a fluorescent aptasensor based on graphene oxide (GO) is developed for the detection of tomato systemin (TomSys) with aptamer recognition properties. In the absence of TomSys, the aptamer-functionalized QDs are adsorbed on the surface of GO and the fluorescence is efficiently quenched, while in the presence of TomSys, the specific binding of TomSys with its aptamer competitively releases aptamer-functionalized QDs from the GO surface, leading to the recovery of QDs fluorescence. The results demonstrate that the simple, rapid and cost-efficient biosensor possesses satisfactory sensitivity and selectivity for the detection of TomSys.

One-Step Synthesis of Rox-DNA Functionalized CdZnTeS Quantum Dots for the Visual Detection of Hydrogen Peroxide and Blood Glucose

As the blood glucose concentration is an important clinical parameter of diabetes, the rapid and effective detection of blood glucose is very significant for monitoring and managing diabetes. Here, a facile method to prepare Rox-DNA functionalized CdZnTeS quantum dots (QDs) was developed. The Rox-DNA functionalized CdZnTeS QDs were prepared by a one-pot hydrothermal method through phosphorothioate DNA bound to QDs, which were employed as a ratiometric fluorescent probe for the rapid and sensitive detection of H2O2 and glucose. Compared with the traditional multistep construction of ratiometric fluorescent probes, this presented approach is simpler and more effective without chemical modification and complicated separation. The CdZnTeS QDs with green fluorescence is specifically sensitive to H2O2, while the red fluorescence of Rox is invariable. H2O2 is the product from the oxidation of glucose catalyzed by glucose oxidase (GOx). Therefore, a facile method to detect H2O2 and glucose with a detection limit of 0.075 μM for H2O2 and 0.042 μM for glucose was developed. In addition, this proposed probe has been employed for the detection of glucose in human serum with a satisfactory result. Moreover, this probe has been used for visual detection, and the health and diabetics can be distinguished by the naked eye. Meanwhile, this nanoprobe is also generalizable and can be extended to the detection of many other H2O2-mediated analytes.

Target-induced structure switching of a hairpin aptamer for the fluorescence detection of zeatin

Zeatin (ZT) is a type of cytokinin which plays important roles in the proliferation and differentiation of plants cells. Here, we describe a fluorometric method for the detection of ZT, based on target-induced structure switching of a hairpin aptamer. The signal probe, an FAM-labeled aptamer, hybridizes partially with BHQ-2-labeled DNA and the fluorescence intensity of FAM is quenched by the approaching BHQ-2. With the addition of the target, the aptamer binds to ZT with high specificity and the BHQ-2-labeled DNA is released. Thus, the fluorescence intensity of the FAM-labeled aptamer is recovered and ZT can be quantitatively determined by monitoring the fluorescence signal change. This proposed method shows good sensitivity with a detection limit as low as 135 nM for ZT detection. In comparison to the previously reported ZT detection strategies based on graphene oxide (GO), this biosensor exhibits excellent selectivity. Moreover, this method has great potential for parallel analysis of different aptamer-based target molecules.

A nonenzymatic DNA nanomachine for biomolecular detection by target recycling of hairpin DNA cascade amplification

Synthetic enzyme-free DNA nanomachine performs quasi-mechanical movements in response to external intervention, suggesting the promise of constructing sensitive and specific biosensors. Herein, a smart DNA nanomachine biosensor for biomolecule (such as nucleic acid, thrombin and adenosine) detection is developed by target-assisted enzyme-free hairpin DNA cascade amplifier. The whole DNA nanomachine system is constructed on gold nanoparticle which decorated with hundreds of locked hairpin substrate strands serving as DNA tracks, and the DNA nanomachine could be activated by target molecule toehold-mediated exchange on gold nanoparticle surface, resulted in the fluorescence recovery of fluorophore. The process is repeated so that each copy of the target can open multiplex fluorophore-labeled hairpin substrate strands, resulted in amplification of the fluorescence signal. Compared with the conventional biosensors of catalytic hairpin assembly (CHA) without substrate in solution, the DNA nanomachine could generate 2-3 orders of magnitude higher fluorescence signal. Furthermore, the DNA nanomachine could be used for nucleic acid, thrombin and adenosine highly sensitive specific detection based on isothermal, and homogeneous hairpin DNA cascade signal amplification in both buffer and a complicated biomatrix, and this kind of DNA nanomachine could be efficiently applied in the field of biomedical analysis.

One-Pot Synthesis of the Stable CdZnTeS Quantum Dots for the Rapid and Sensitive Detection of Copper-Activated Enzyme

Galactose oxidase is a copper-activated enzyme and have a vital role in metabolism of galactose. Much of the work is focused on determining the amount of galactose in the blood rather than measuring the amount of galactose oxidase to urge the galactosemia patients to restrict milk intake. Here, a simple and effective method was developed for Cu2+ and copper-activated enzyme detection based on homogenous alloyed CdZnTeS quantum dots (QDs). Meso- 2,3-dimercaptosuccinic acid (DMSA) was used as the reducing agent for preparing QDs and the highest quantum yield of CdZnTeS QDs was 69.4%. In addition, the as-prepared CdZnTeS QDs show superior fluorescence properties, such as good photo-/chemical stability. The DMSA was the surface ligand of the QDs, containing abundant -SH and -COOH, thus the surface ligands have a high affinity with Cu2+. Therefore, this developed probe can be applied for Cu2+ and galactose oxidase detection and shows a good sensitivity in the buffer. Then, this probe was successfully used for Cu2+ and galactose oxidase detection in real samples with the satisfactory results. The proposed fluorescence quenching strategy gives a new and simple insight for enzyme assay without the enzyme-catalyzed reaction.

The synthesis of a smart streptavidin-functionalized poly(N-isopropylacrylamide) composite and its application in the separation and detection of virus nucleic acid

A new kind of polymeric material (PNIPAAm-co-SA) was prepared by conjugating a thermosensitive polymer, Poly (N-isopropylacrylamide) (PNIPAAm) with streptavidin (SA). This smart prepared composite displayed a controllable conformation change between an expanded and a collapsed form, below or above its lower critical solution temperature (LCST). Differential scanning calorimetry (DSC) analysis demonstrated that the PNIPAAm-co-SA bioconjugate showed the same LCST as the original synthetic polymer, PNIPAAm, which was also 32°C. Based on the specific interaction between SA and biotin, a higher capture efficiency of PNIPAAm-co-SA, which was almost 100% in PBS buffer solution and above 70% in serum was obtained, respectively. And the high affinity between PNIPAAm-co-SA and biotin was still maintained after three heating cycles. Subsequently, the variola virus (small pox, VV) oligonucleotide sequence was chosen as a model to demonstrate the sensitivity of the biosensor which was fabricated based on PNIPAAm-co-SA. The biosensor exhibited the ability to separate and enrich targets from complicated system with its phase transition ability, and high sensitivity toward VV-targets were achieved. Moreover, other types of targets such as proteins and cells, could be detected by changing the biotin-captures, which indicated the broad applicability of biosensors based on this smart polymer material.

Rox-DNA Functionalized Silicon Nanodots for Ratiometric Detection of Mercury Ions in Live Cells

A ratiometric fluorescent sensor for mercury ions (Hg2+) has been constructed via covalent functionalization of silicon nanodot (SiND) with Hg2+-specific 6-carboxy-X-rhodamine (Rox)-tagged DNA. For the Rox-DNA functionalized SiND, the red fluorescence of Rox can be quenched by the blue-emitting SiND in the presence of Hg2+ due to structural change in DNA, which serves as the response signal. Meawhile, the fluorescence of SiND is insensitive to Hg2+ and acts as the reference signal. The wavelength difference in the optimal emission peak is as large as 190 nm between SiND (422 nm) and Rox (612 nm), which can efficaciously exclude the interference of the two emission peaks, and facilitates dual-color visualization of Hg2+ ions. The biofunctionalization of SiND improves the acid-base stability of SiND significantly, which is favorable for its application in the intracellular environment. Accordingly, a sensitive, simple, precise and rapid method for tracing Hg2+ was proposed. The limit of detection and precision of this method for Hg2+ was 9.2 nM and 8.8% (50 nM, n = 7), respectively. The increase of Hg2+ concentration in the range of 10-1500 nM was in accordance with linearly increase of the I422/ I612 ratio. As for practical application, the recoveries in spiked human urine and serum samples were in the range of 81-107%. Moreover, this fluorescent nanosensor was utilized to the ratiometric detection of Hg2+ in HeLa cells.

Silicon nanodot-based aptasensor for fluorescence turn-on detection of mucin 1 and targeted cancer cell imaging

We report herein a new dual-color fluorescent aptasensor for detection of tumor marker mucin 1 (MUC1) and targeted imaging of MCF-7 cancer cells based on the specific interaction between MUC1 and its aptamer S2.2. The aptasensor was prepared by covalent attachment of the cyanine (Cy5)-tagged aptamer S2.2 to fluorescent silicon nanodot (SiND). The fluorescence of S2.2-Cy5 could be quenched by the SiND carrier in the absence of MUC1, and its fluorescence was restored in the presence of MUC1 due to structure switching of S2.2. This aptasensor exhibits specificity for MUC1-possitive MCF-7 cells rather than MUC1-negative MCF-10A cells and Vero cells. The SiND plays multiple roles in this fluorescence assay, making the method easier compared with other approaches. The limit of detection and precision of this method for MUC1 was 1.52 nM and 3.6% (10 nM, n = 7), respectively. The linear range was 3.33-250 nM, and the recoveries in spiked human serum were in the range of 87-108%. This is a simple, selective, sensitive and reliable method, which can well achieve not only quantitative analysis of tumor marker but also dual-color visualization of single cancer cells.

Facile synthesis of stable CdTe/CdS QDs using dithiol as surface ligand for alkaline phosphatase detection based on inner filter effect

Alkaline phosphatase (ALP) is a universal and important hydrolase that has been proved to be associated with several diseases. Herein, a simple and effective method was proposed for ALP detection based on the inner filter effect of p-nitrophenol (pNP) on the fluorescence of CdTe/CdS quantum dots (QDs). For the preparation of CdTe/CdS QDs, Na2TeO3 was used as the Te source, and dithiol as the S source and surface ligand. The as-prepared CdTe/CdS QDs show good fluorescence properties, such as high quantum yield (∼80%), and good chemical/photo-stability. pNP is a hydrolysate of p-nitrophenol phosphate disodium salt under the catalysis of ALP, which could effectively quench the fluorescence of QDs due to the absorption spectra of pNP overlaps well with the excitation spectra of the CdTe/CdS QDs. Therefore, the prepared CdTe/CdS QDs could be applied for ALP detection. A good linear relationship ranging from 2.2 to 220 U/L was obtained with the limit of detection as low as 0.34 U/L. In addition, this method was successfully applied for the assay of ALP in human serum with the satisfactory results.