Bingling Li

Simple and sensitive aptamer-based colorimetric sensing of protein using unmodified gold nanoparticle probes

We describe herein simple and sensitive aptamer-based colorimetric sensing of protein (alpha-thrombin in this work) using unmodified gold nanoparticle probes.

Highly ordered mesoporous carbons as electrode material for the construction of electrochemical dehydrogenase- and oxidase-based biosensors

In this work, the excellent catalytic activity of highly ordered mesoporous carbons (OMCs) to the electrooxidation of nicotinamide adenine dinucleotide (NADH) and hydrogen peroxide (H(2)O(2)) was described for the construction of electrochemical alcohol dehydrogenase (ADH) and glucose oxidase (GOD)-based biosensors. The high density of edge-plane-like defective sites and high specific surface area of OMCs could be responsible for the electrocatalytic behavior at OMCs modified glassy carbon electrode (OMCs/GE), which induced a substantial decrease in the overpotential of NADH and H(2)O(2) oxidation reaction compared to carbon nanotubes modified glassy carbon electrode (CNTs/GE). Such ability of OMCs permits effective low-potential amperometric biosensing of ethanol and glucose, respectively, at Nafion/ADH-OMCs/GE and Nafion/GOD-OMCs/GE. Especially, as an amperometric glucose biosensor, Nafion/GOD-OMCs/GE showed large determination range (500-15,000 micromoll(-1)), high sensitivity (0.053 nA micromol(-1)), fast (9+/-1s) and stable response (amperometric response retained 90% of the initial activity after 10h stirring of 2 mmoll(-1) glucose solution) to glucose as well as the effective discrimination to the possible interferences, which may make it to readily satisfy the need for the routine clinical diagnosis of diabetes. By comparing the electrochemical performance of OMCs with that of CNTs as electrode material for the construction of ADH- and GOD-biosensors in this work, we reveal that OMCs could be a favorable and promising carbon electrode material for constructing other electrochemical dehydrogenase- and oxidase-based biosensors, which may have wide potential applications in biocatalysis, bioelectronics and biofuel cells.

Sensitive detection of protein by an aptamer-based label-free fluorescing molecular switch

We report an aptamer-based method for the sensitive detection of proteins by a label-free fluorescing molecular switch (ethidium bromide), which shows promising potential in making protein assay simple and economical.

DNA based gold nanoparticles colorimetric sensors for sensitive and selective detection of Ag(I) ions

In this work, we reported both unlabeled and labeled sensing strategies for Ag(I) ions detection by using the DNA based gold nanoparticles (AuNPs) colorimetric method. In the unlabeled strategy, C-base riched single strand DNA (C-ssDNA) enwinded onto AuNPs to form AuNPs/C-ssDNA complex. In the labeled method, sulfhydryl group modified C-ssDNA (HS-C-ssDNA) was covalently labeled on AuNPs to produce AuNPs-S-C-ssDNA complex. In both strategies, C-ssDNA or HS-C-ssDNA could enhance the AuNPs stability against the salt-induced aggregation. However, the presence of Ag(I) ions in the obtained AuNPs/C-ssDNA or AuNPs-S-C-ssDNA complex would decrease such stability to display purple even blue colors due to the formation of Ag(I) ions mediated C-Ag(I)-C base pairs. Through this phenomenon, Ag(I) ions could be detected qualitatively and quantitatively using both unlabeled and labeled sensing strategies. Compared with the labeled method, the unlabeled strategy avoided the label and separation steps in common sensors, which may thus save both the time and cost for the detection. Nevertheless, the labeled strategy provided more sensitive, stable and controllable sensing results compared with the unlabeled method. By the labeled strategy, 12 nM Ag(I) ions could be observed directly by naked eyes, and the lowest detectable concentration of 0.59 nM was gotten under by the UV-vis spectra measurement, which was one of the most sensitive results among DNA based AuNPs colorimetric sensors for metal ions.

Au NPs-enhanced surface plasmon resonance for sensitive detection of mercury(II) ions.

We reported a sensitive surface plasmon resonance (SPR) sensor for the detection of Hg(2+) in aqueous solution by using a thymine (T)-rich, mercury-specific oligonucleotide (MSO) probe and gold nanoparticles (Au NPs)-based signal amplification. The MSO probe was first immobilized on gold film through formation of Au-S bond between DNA and gold film. In the presence of Hg(2+), the MSO probe captured free Hg(2+) in aqueous media via the Hg(2+)-mediated coordination of T-Hg(2+)-T base pairs. This direct immobilization strategy led to a detection limit of 0.3 microM of Hg(2+). In order to improve the sensitivity, part complementary DNA (PCS)-modified Au NPs labels were employed to amplify SPR signals. We demonstrated that this Au NPs-based sensing strategy resulted in a detection limit down to 5 nM of Hg(2+), brings about an amplification factor of two orders of magnitude. This Au NPs-based Hg(2+) sensor also exhibited excellent selectivity over a spectrum of interference metal ions. Taking advantage of the high amplifying characteristic of Au NPs and the specificity of MSO to Hg(2+) recognition, we developed here a SPR sensor for specific Hg(2+) detection with high sensitivity.

Layer-by-layer electrochemical biosensor with aptamer-appended active polyelectrolyte multilayer for sensitive protein determination

Herein, we report two simple label-free electrochemical aptasensors for protein detection using layer-by-layer (LBL) self-assembled multilayers with ferrocene-appended poly(ethyleneimine) (Fc-PEI), carbon nanotubes (CNTs) and aptamer. In one sensing strategy, the Fc-PEI, CNTs and DNA aptamer are LBL assembled on the electrode surface via electrostatic interaction. In the presence of target, the aptamer on the outermost layer of the LBL self-assembled multilayer would catch the target on the electrode interface, which makes a barrier for electrons and inhibits the electro-transfer, resulting in the decreased DPV signals of Fc-PEI. Using this strategy, a wide detection range (0.3-165 ng ml(-1)) for model target thrombin is obtained, with a low detection limit of 0.14 ng ml(-1). In the similar sensing strategy for detection of lysozyme, a wide detection range (0.2 ng ml(-1) to 1.66 microg ml(-1)) and a low detection limit (0.17 ng ml(-1)) are obtained. These results prove that the LBL sensing strategies developed possess sensitivity, selectivity, stability and generality.

DNA G-quadruplex-templated formation of the fluorescent silver nanocluster and its application to bioimaging.

Herein, a novel kind of silver nanocluster is synthesized simply by mixing G-quadruplex template with silver ions and reduction reagent (NaBH(4), here). AS1411 (a G-quadruplex that can bind nucleolin overexpressed in cancer cells) is used as the main model template to prove the synthesis protocol and its potential application. We used fluorescence assay, CD, MALDI TOF MS, and TEM to characterize the silver nanocluster. It is found that after formation of the silver nanocluster, AS1411 still keeps its structure and is able to bind with nucleolin in cancer cell. Meanwhile, this binding behavior can greatly enhance the fluorescence intensity of the silver nanocluster. This property can be directly employed into bioimaging HeLa cells. The cell toxicity (3-[4,5-dimethylthiazolyl-2]-2,5-diphenyltetrazolium bromide, MTT) assay demonstrated that the silver nanocluster has only little affect on the cytotoxicity to the cells, which further proves the applicability of the method in tumor cell imaging. At last, the universality of the synthesis protocol is verified by using a series of other G-quadruplex sequences as templates. For a lot of functional nucleic acids, such as human telomeres and certain aptamers, are with G-rich sequences and can fold into G-quadruplexes in functioning conditions, our method displays a promising application space in future researches.

[Ru(bpy)(2)(dcbpy)NHS] Labeling/Aptamer-Based Biosensor for the Detection of Lysozyme by Increasing Sensitivity with Gold Nanoparticle Amplification

A novel [Ru(bpy)(2)(dcbpy)NHS] labeling/aptamer-based biosensor combined with gold nanoparticle amplification for the determination of lysozyme with an electrochemiluminescence (ECL) method is presented. In this work, an aptamer, an ECL probe, gold nanoparticle amplification, and competition assay are the main protocols employed in ECL detection. With all the protocols used, an original biosensor coupled with an aptamer and [Ru(bpy)(2)(dcbpy)NHS] has been prepared. Its high selectivity and sensitivity are the main advantages over other traditional [Ru(bpy)(3)](2+) biosensors. The electrochemical impedance spectroscopy (EIS) and atomic force microscopy (AFM) characterization illustrate that this biosensor is fabricated successfully. Finally, the biosensor was applied to a displacement assay in different concentrations of lysozyme solution, and an ultrasensitive ECL signal was obtained. The ECL intensity decreased proportionally to the lysozyme concentration over the range 1.0x10(-13)-1.0x10(-8) mol L(-1) with a detection limit of 1.0x10(-13) mol L(-1). This strategy for the aptasensor opens a rapid, selective, and sensitive route for the detection of lysozyme and potentially other proteins.

Reusable, label-free electrochemical aptasensor for sensitive detection of small molecules

We report a sensitive electrochemical aptasensor for adenosine based on electrochemical impedance spectroscopy measurement, which gives not only a label-free but also a reusable platform to make the detection of small molecules simple and convenient.

In situ labeling and imaging of cellular protein via a bi-functional anticancer aptamer and its fluorescent ligand

In this article, we reported a novel approach for in situ labeling and imaging HeLa cancer cells utilizing a bifunctional aptamer (AS1411) and its fluorescent ligand, protoporphyrin IX (PPIX). In the presence of potassium ion, AS1411 folded to G-quadruplex structure, binded fluorescent ligand (PPIX) with fluorescent enhancement, and targeted the nucleolin overexpressed by cancer cells. Consequently, bioimaging of cancer cells specifically were realized by laser scanning confocal microscope. The bioimaging strategy with AS1411-PPIX complex was capable to distinguish HeLa cancer cells from normal cells unambiguously, and fluorescence imaging of cancer cells was also realized in human serum. Moreover, the bioimaging method was very facile, effective and need not any covalent modification. These results illustrated that the useful approach can provide a novel clue for bioimaging based on non-covalent bifunctional aptamer in clinic diagnosis.