Chih-Ching Huang

Detection of mercury(II) based on Hg2+–DNA complexes inducing the aggregation of gold nanoparticles

A DNA-Au NP probe for sensing Hg2+ using the formation of DNA-Hg2+ complexes through thymidine (T)-Hg2+ -T coordination to control the negative charge density of the DNA strands-thereby varying their structures-adsorbed onto Au NPs.

Oligonucleotide-Based Fluorescence Probe for Sensitive and Selective Detection of Mercury(II) in Aqueous Solution

In this paper we unveil a new homogeneous assayusing TOTO-3 and the polythymine oligonucleotide T 33for the highly selective and sensitive detection of Hg (2+) in aqueous solution. The fluorescence of TOTO-3 is weak in the absence or presence of randomly coiled T 33. After T 33 interacts specifically with Hg (2+) ions through T-Hg (2+)-T bonding, however, its conformation changes to form a folded structure that preferably binds to TOTO-3. As a result, the fluorescence of a mixture of T 33 and TOTO-3 increases in the presence of Hg (2+). Our data from fluorescence polarization spectroscopy, capillary electrophoresis with laser-induced fluorescence detection, circular dichroism spectroscopy, and melting temperature measurements confirm the formation of folded T 33-Hg (2+) complexes. Under optimum conditions, the TOTO-3/T 33 probe exhibited a high selectivity (>or=265-fold) toward Hg (2+) over other metal ions, with a limit of detection of 0.6 ppb. We demonstrate the practicality of this TOTO-3/T 33 probe for the rapid determination of Hg (2+) levels in pond water and in batteries. This approach offers several advantages, including rapidity (<15 min), simplicity (label-free), and low cost.

Parameters for selective colorimetric sensing of mercury(II) in aqueous solutions using mercaptopropionic acid-modified gold nanoparticles

We unveil a new homogeneous assay-using mercaptopropionic acid-modified Au nanoparticles in the presence of 2,6-pyridinedicarboxylic acid for the highly selective and sensitive detection of Hg(2+) ions.

Synthesis of Fluorescent Carbohydrate-Protected Au Nanodots for Detection of Concanavalin A and Escherichia coli

This study describes a novel, simple, and convenient method for the preparation of water-soluble biofunctional Au nanodots (Au NDs) for the detection of Concanavalin A (Con A) and Escherichia coli (E. coli). First, 2.9 nm Au nanoparticles (Au NPs) were prepared through reduction of HAuCl(4).3H(2)O with tetrakis(hydroxymethyl)phosphonium chloride (THPC), which acts as both a reducing and capping agent. Addition of 11-mercapto-3,6,9-trioxaundecyl-alpha-D-mannopyranoside (Man-SH) onto the surfaces of the as-prepared Au NPs yielded the fluorescent mannose-protected Au nanodots (Man-Au NDs) with the size and quantum yield (QY) of 1.8 (+/-0.3) nm and 8.6%, respectively. This QY is higher than those of the best currently available water-soluble, alkanethiol-protected Au nanoclusters. Our fluorescent Man-Au NDs are easily purified and by multivalent interactions are capable of sensing, under optimal conditions, Con A with high sensitivity (LOD = 75 pM) and remarkable selectivity over other proteins and lectins. To the best of our knowledge, this approach provided the lowest LOD value for Con A when compared to the other nanomaterials-based detecting method. Furthermore, we have also developed a new method for fluorescence detection of E. coli using these water-soluble Man-Au NDs. Incubation with E. coli revealed that the Man-Au NDs bind to the bacteria, yielding brightly fluorescent cell clusters. The relationship between the fluorescence signal and the E. coli concentration was linear from 1.00 x 10(6) to 5.00 x 10(7) cells/mL (R(2) = 0.96), with the LOD of E. coli being 7.20 x 10(5) cells/mL.

Bioconjugated gold nanodots and nanoparticles for protein assays based on photoluminescence quenching.

This study describes the first instance of the use of two differently sized Au nanoparticles (Au NPs), acting separately as donor and acceptor, in homogeneous photoluminescence quenching assays developed for the analysis of proteins. Introduction of a breast cancer marker protein, platelet-derived growth factor AA (PDGF AA), to a solution of 11-mercaptoundecanoic acid-protected, 2.0-nm photoluminescent Au nanodots (L(AuND)) led to the preparation of PDGF AA-L(AuND) as the donor. Thiol-derivative PDGF binding aptamers (Apt) and 13-nm spherical Au NPs were used to synthesize the Apt-Q(AuNP) acceptor. The photoluminescence of PDGF AA-L(AuND) at 520 nm decreased when photoluminescence quenching occurred between Apt-Q(AuNP) and PDGF AA-L(AuND). We used the PDGF AA-L(AuND)/Apt-Q(AuNP)-based molecular light switching system to analyze PDGFs and PDGF alpha-receptor in separate homogeneous solutions. In the presence of PDGFs, the interaction between Apt-Q(AuNP) and PDGF AA-L(AuND) decreased as a result of competitive reactions between the PDGFs and Apt-Q(AuNP). Similarly, the interaction between Apt-Q(AuNP) and PDGF AA-L(AuND) reduced as a result of competitive reactions between PDGF alpha-receptor and PDGF AA-L(AuND). The limits of detection (LODs) for PDGF AA and PDGF alpha-receptor were 80 pM and 0.25 nM, respectively, resulting from a low background photoluminescence signal. When using the Apt-Q(AuNP) as selectors for (a) the enrichment of PDGF AA and (b) the removal of matrixes possessing intense background fluorescence from cell media and urine samples, the LOD for PDGF AA decreased to 10 pM. Unlike quantum dots, the L(AuND) provide the advantages of biocompatibility, ease of bioconjugation, and minimal toxicity.

Colorimetric Assay for Lead Ions Based on the Leaching of Gold Nanoparticles

A colorimetric, label-free, and nonaggregation-based gold nanoparticles (Au NPs) probe has been developed for the detection of Pb(2+) in aqueous solution, based on the fact that Pb(2+) ions accelerate the leaching rate of Au NPs by thiosulfate (S(2)O(3)(2-)) and 2-mercaptoethanol (2-ME). Au NPs reacted with S(2)O(3)(2-) ions in solution to form Au(S(2)O(3))(2)(3-) complexes on the Au NP surfaces, leading to slight decreases in their surface plasmon resonance (SPR) absorption. Surface-assisted laser desorption/ionization time-of-flight ionization mass spectrometry (SALDI-TOF MS) data reveals the formation of Pb-Au alloys on the surfaces of the Au NPs in the presence of Pb(2+) ions and 2-ME. The formation of Pb-Au alloys accelerated the Au NPs rapidly dissolved into solution, leading to dramatic decreases in the SPR absorption. The 2-ME/S(2)O(3)(2-)-Au NP probe is highly sensitive (LOD = 0.5 nM) and selective (by at least 1000-fold over other metal ions) toward Pb(2+) ions, with a linear detection range (2.5 nM-10 muM) over nearly 4 orders of magnitude. The cost-effective probe allows rapid and simple determination of the concentrations of Pb(2+) ions in environmental samples (Montana soil and river), with results showing its great practicality for the detection of lead in real samples.

Colorimetric determination of urinary adenosine using aptamer-modified gold nanoparticles

This paper describes a colorimetric sensing approach for the determination of adenosine triphosphate (ATP) using aptamer-modified gold nanoparticles (Apt-Au NPs). In the absence of the analytes, the color of the Apt-Au NPs solution changed from wine-red to purple as a result of salt-induced aggregation. Binding of the analytes to the Apt-Au NPs induced folding of the aptamers on the Au NP surfaces into four-stranded tetraplex structures (G-quartet) and/or an increase in charge density. As a result, the Apt-Au NPs solution was wine-red in color in the presence of the analytes under high salt conditions. For mixtures of ATP (20.0-100.0nM), Apt-Au NPs (3.0nM), 10.0% poly(ethylene glycol), 0.2microM TOTO-3, 150.0mM NaCl, 15.0mM KCl, and 16.0mM Tris-HCl (pH 7.4), a linear correlation (R(2)=0.99) existed between the ratio of the extinctions of the Apt-Au NPs at 650 and 520nm (Ex(650/520)) and the concentration of ATP. The limit of detection for ATP was 10.0nM. The practicality of this simple, sensitive, specific, and cost-effective approach was demonstrated through the determination of the concentration of adenosine in urine samples.

Control over surface DNA density on gold nanoparticles allows selective and sensitive detection of mercury(II).

We have developed a new highly selective and sensitive technique for the detection of Hg(2+) using DNA-functionalized gold nanoparticles (Au NPs) and OliGreen. This system is the first that allows the detection of Hg(2+) based on the release of DNA molecules, induced by conformational changes on Au NP surfaces, and its sensitivity is highly dependent upon surface DNA density. When Hg(2+) ions interact with the thymidine units of the DNA molecules bound to the Au NPs through Au-S bonds, the conformations of these DNA derivatives change from linear to hairpin structures, causing the release of some of the DNA molecules from the surface of the Au NPs into the bulk solution to react with OliGreen. The fluorescence of OliGreen-DNA complexes increased with increasing concentration of Hg(2+), and Hg(2+) could be detected at concentrations as low as 25 nM. A linear correlation existed between the fluorescence intensity and the concentration of Hg(2+) over the range 0.05-2.5 microM (R(2) = 0.98). This simple and cost-effective probe was applied to determine the spiked Hg(2+) in the pond samples; the recoveries (96-102%) suggested low matrix interference and high sensitivity.

Fluorescent gold and silver nanoclusters for the analysis of biopolymers and cell imaging

Fluorescent gold and silver nanoclusters are interesting sensing materials because of their molecule-like optical properties, easy preparation, and biocompatibility. In this review, we highlight the chemical and optical properties of fluorescent gold and silver nanoclusters, as well as their preparation and applications in biomolecular analysis and cell imaging.

Label-free colorimetric detection of picomolar thrombin in blood plasma using a gold nanoparticle-based assay.

We unveil a novel, label-free, colorimetric assay--using fibrinogen (Fib) and gold nanoparticles (Au NPs)--for the highly selective and sensitive detection of thrombin. Addition of fibrinogen to a solution of Au NPs (average diameter: 56 nm) led to ready conjugation, forming Fib-Au NPs through electrostatic and hydrophobic interactions. Introduction of thrombin (a serine protease) into the Fib-Au NPs solutions in the presence of excess fibrinogen induced the formation of insoluble fibrillar fibrin-Au NPs agglutinates through the polymerization of the unconjugated and conjugated fibrinogen. After centrifugation, the absorbance at 532 nm of the supernatants decreased upon increasing the concentration of thrombin. This Fib-Au NP probe provides high sensitivity [limit of detection (LOD): 0.04 pM] for thrombin, with remarkable selectivity over other proteins and proteases. The range of linearity for the absorbance against the thrombin concentration was 0.1-10 pM (R(2)=0.96). This approach provides an LOD for thrombin that is lower than those obtainable using other nanomaterial- and aptamer-based detection methods. We validated the utility of this Fib-Au NP probe through separate analyses of thrombin and Factor Xa at picomolar levels in plasma samples--without the need for sample pretreatment. This technique appears to have practical potential in the diagnosis of diseases associated with coagulation abnormalities and cancers (e.g., pulmonary metastasis).