An aptamer based method is described for the electrochemical determination of ampicillin. It is based on the use of DNA aptamer, DNA functionalized gold nanoparticles (DNA-AuNPs), and single-stranded DNA binding protein (ssDNA-BP). When the aptamer hybridizes with the target DNA on the AuNPs, the ssDNA-BP is captured on the electrode surface via its specific interaction with ss-DNA. This results in a decreased electrochemical signal of the redox probe Fe(CN)63? which is measured best at a voltage of 0.188 mV (vs. reference electrode). In the presence of ampicillin, the formation of aptamer-ampicillin conjugate blocks the further immobilization of DNA-AuNPs and ssDNA-BP, and this leads to an increased response. The method has a linear reposne that convers the 1 pM to 5 nM ampicillin concentration range, with a 0.38 pM detection limit (at an S/N ratio of 3). The assay is selective, stable and reproducible. It was applied to the determination of ampicillin in spiked milk samples where it gave recoveries ranging from 95.5 to 105.5%.
Graphical abstract Schematic of a simple and sensitive electrochemical apta-biosensor for ampicillin detection. It is based on the use of gold nanoparticles (AuNPs), DNA aptamer, DNA functionalized AuNPs (DNA-AuNPs), and single-strand DNA binding protein (SSBP).
A study is presented on the binding kinetics and mechanism of the adsorption of dsDNA on citrate-capped gold nanoparticles (AuNPs). Methods include fluorescence titration, isothermal calorimetry (ITC) titration, dynamic light scattering and gel electrophoresis. It is found that the fluorescence of probe DNA (labeled with Rhodamine Green and measured at excitation/emission peaks of 498/531 nm) is quenched by addition of AuNPs. The Stern-Volmer quenching constant (Ksv) is 1.67?×?10^9 L·mol?1 at 308 K and drops with increasing temperature. The quenching mechanism is mainly static. The results of both fluorescence titrations and ITC show negative values for ΔH and ΔS values. This shows ion-induced dipole-dipole interaction to be the main attractive forces between dsDNA and AuNPs, while electrostatic interactions result in repulsion. The repulsive forces lead to a lower affinity between dsDNA and AuNPs (compared to single-strand DNA). It is also found that dsDNA can prevent the aggregation of AuNPs which is accompanied by a color change from red into blue. The visual detection limit with bare eyes for dsDNA1 is 36 pM. Based on these findings, a colorimetric method was developed to detect the proto-oncogene of serine/threonine-protein kinase B-Raf V600E point mutation in HT29, Ec109, A549, Huh-7 and SW480 cell lines.
Graphical abstract Schematic of the salt-induced aggregation of uncapped gold nanoparticles (AuNPs) which leads to a color change from red to blue. If the AuNPs are coated with dsDNA, aggregation is suppressed.
The authors describe a colorimetric method for the determination of DNA based on the deaggregation of gold nanoparticles (AuNPs) induced by exonuclease III (Exo III). DNA amplification is accomplished by Exo III to generate large quantities of the residual DNA. Residual DNA tethers onto the surfaces of AuNPs which prevents their aggregation. Hence, the color of the solution is red. However, in the absence of DNA, salt-induced aggregation is not prevented, and the bluish-purple color of the aggregated AuNPs is observed. The ratio of absorbances at 525 and 625 nm increases up to 150 nM DNA concentrations, and the LOD is as low as 3.0 nM. It is shown that the presence of 300 nM concentrations of random DNA (with a mass up to 10-fold that of target DNA) does not interfere. The method was successfully applied to the analysis of DNA in spiked serum samples. The method is simple, reliable, and does not require complicated amplification steps and expensive instrumentation.
Graphical abstract Schematic of a sensing strategy for DNA detection by exonuclease III-induced deaggregation of gold nanoparticles. DNA concentrations as low as 3 nM can be detected via colorimetric monitoring of the color change from red to purple-blue.
The paper describes a voltammetric method for the quantitation of the activity of telomerase extracted from cancer cells. A thiolated single-stranded telomerase substrate primer was firstly immobilized on a gold electrode. In the presence of a mixture of telomerase and deoxynucleotide triphosphates, the primer becomes elongated and contains repetitive nucleotide sequences (TTAGGG)n. After hybridization with blocker DNA, gold nanoparticles are added and captured by the elongated single-stranded DNA. This reduces the charge transfer resistance of the gold electrode. The telomerase activity is then quantified via differential pulse voltammetry, typically at 0.12 V (vs. SCE). The method is PCR-free, rapid, and convenient. It was applied to the detection of HeLa cells via the telomerase activity of lysed cells. The detection range was from 500 to 50,000 cells/mL and the detection limit was as low as 500 cells/mL.
Graphical abstract A telomerase substrate (TS) primer is immobilized on a gold electrode as the sensing interface to detect the activity of telomerase extracted from cancer cells. Unmodified gold nanoparticles (AuNPs) are utilized which change the electrochemical responses.
The authors describe a voltammetric immunosensor with antibody immobilized on a glassy carbon electrode (GCE) modified with N-doped graphene (N-GS), electrodeposited gold nanoparticles (AuNPs) and chitosan (Chit). The preparation is simple and the thickness of the electrodeposited films can be well controlled. Due to the specific advantages of N-GS, AuNPs and Chit, the electrode has a large specific surface, improved conductivity, high stability. A new label-free immunosensor for the model antigen (alpha fetoprotein, AFP) detection was then designed by employing N-GS-AuNP-Chit as the antibody immobilization and signal amplification platform. Differential pulse voltammetry and electrochemical impedance spectroscopy were used for the characterization of the stepwise assembly process. Under the optimized conditions, at a typical working potential of +0.20 V (vs. SCE), and by using hexacyanoferrate as an electrochemical probe, the immunosensor has a detection limit as low as 1.6 pg mL?1 and a linear analytical range that extends from 5 pg mL?1 to 50 ng mL?1. AFP was quantified in spiked human serum samples with acceptable precision.
Graphical Abstract Schematic of sensitive and effective label-free electrochemical immunosensor for the detection of AFP based on N-GS-AuNP-Chit as signal amplification matrix.
The authors describe a fluorescence based aptasensor for adenosine (AD), a conceivable biomarker for cancer. The assay is based on the immobilization of capture DNA on newly synthesized quaternary CuInZnS quantum dots (QDs) and the conjugation of probe DNA on gold nanoparticles (AuNPs). The capture DNA is an adenosine-specific aptamer that is partly complementary to the probe DNA. Once the capture aptamer hybridizes probe DNA, the fluorescence of the QDs (measured at excitation/emission wavelengths of 522/650 nm) is quenched by the AuNPs. However, when AD is added, it will bind to the aptamer and restrain the hybridization between capture DNA and probe DNA. Therefore, the fluorescence of the QDs will increase with increasing AD concentration. Under optimal conditions, fluorescence is linearly related to the AD concentration in the range from 50 to 400 μM, the detection limit being 1.1 μM. This assay is sensitive, selective, reproducible and acceptably stable. It was applied to the determination of AD in spiked human serum samples where it gave satisfactory results.
The authors describe a gold nanoparticle (AuNP) based aggregation assay for colorimetric determination of silver ions. The detection scheme is based on the release of aptamers from the surface of AuNPs that is triggered by the formation of C-Ag(I)-C links. In the absence of Ag(I) ions, the aptamers are readily adsorbed on the surface of the AuNPs. This prevents the aggregation of AuNPs and warrants the stability of the red colloidal solution at high salt concentration. In the presence of Ag(I) ions, the aptamers are released from the surface of AuNPs due to binding to Ag(I). Hence, salt-induced aggregation of AuNPs will occur which is accompanied by a gradual color change from red to blue. The color change occurs in the 1 to 500 nM Ag(I) concentration range, and the detection limit is 0.77 nM. The method was successfully applied to the determination of Ag(I) in spiked tap water samples.
Graphical abstract Schematic of a gold nanoparticle-based aggregation assay for colorimetric determination of silver ions. Visual quantitation also is posssible due to a gradual color change from red to blue.
The authors describe a method for functionalization of gold nanoparticles (AuNPs) with the supramolecular host molecule, curcubit[7]uril (CB[7]) which can bind rhodamine B (RhB). The fluorescence of RhB is quenched by the AuNPs via surface energy transfer. On addition of ATP, a dimeric RhB-ATP complex is formed and RhB is pushed out of CB[7]. Hence, fluorescence increases by a factor of 8. This fluorescence recovery effect has been utilized to develop a new detection scheme for ATP. The assay, measured at fluorescence excitation and emission wavelengths of 500 nm and 574 nm respectively, works in the 0.5–10 μM concentration range and has a 100 nM detection limit. The method is not interfered by UTP, GTP, CTP, TTP, ascorbic acid and glutathione.
Graphical abstract Schematic of a method for determination of ATP in the 500 nM to 10 μM concentration range by using fluorescence recovery after surface energy transfer (SET) between rhodamine B (RhB) and gold nanoparticles capped with curcubit[7]uril (CB[7]).
A glassy carbon electrode (GCE) was modified with gold nanoparticles (AuNPs) coated on monolayer graphene (AuNP/MG) by direct in situ sputtering of AuNPs on CVD-generated graphene. This process avoids complicated polymer transfer and polymer cleaning processes and affords AuNPs with a clean surface. The monolayer graphene is ductile and well dispersed. The clean surface of the AuNPs renders this sensor superior to GCEs modified with AuNPs on reduced graphene oxide in terms of the amperometric non-enzymatic determination of hydrogen peroxide. The detection limit is 10 nM (S/N = 3) at 0.55 V (vs. SCE), which is lower than that for similar methods, and the response time is as short as 2 s. Another attractive feature of the sensor is its feasibility for large-scale production via CVD and sputtering.
Graphical abstract Gold nanoparticles deposited onto monolayered graphene generated by chemical vapor deposition (CVD) are used for electrochemical sensing of H2O2, with the detection limit of 10 nM (S/N = 3) and response time of less than 2 s.
The authors describe a method for the fabrication of a nanohybrid composed of carbon dots (C-dots) and gold nanoparticles (AuNPs) by in-situ reduction of C-dots and hydroauric acid under alkaline conditions. The process does not require the presence of surfactant, stabilizing agent, or reducing agent. The hybrid material was deposited in a glassy carbon electrode (GCE), and the modified GCE exhibited good electrocatalytic activity toward the oxidation of nitrite due to the synergistic effects between carbon dots and AuNPs. The findings were used to develop an amperometric sensor for nitrite. The sensor shows a linear response in the concentration range from 0.1 μmol?L-1 to 2 mmol?L-1 and a low detection limit of 0.06 μmol?L-1 at the signal-to-noise ratio of 3.
Graphical abstract Fabrication, characterization and electrochemical behavior of a glassy carbon electrode modifid with carbon dots and gold nanoparticles for sensing nitrite in lake water.
The authors describe an aptasensor for visual and fluorescent detection of lysozyme via an inner filter effect (IFE). The assay is based on the fact that red gold nanoparticles (AuNPs) act as powerful absorbers of the green fluorescence of CdTe because of spectral overlap. If the lysozyme-binding aptamer is adsorbed onto the surface of the AuNPs, the salt-induced aggregation of AuNPs (that leads to a color change from red to blue) does not occur and the IFE remains efficient. If lysozyme is present, it will bind the aptamer and thereby prevent its adsorption on the AuNPs. As a result, the salt-triggered aggregation of the AuNPs will occur. Consequently, color will change from red to blue, and green fluorescence will pop up because the IFE is suppressed. Under optimum conditions, fluorescence is linearly related to lysozyme concentration in the 1.0 nM to 20 nM concentration range, with a 0.55 nM limit of detection. The method is perceived to be of wider applicability in that it may be used to design other visual and fluorescent assays if appropriate aptamers are available.
Graphical abstract The fluorescence intensity of QDs is quenched by gold nanoparticles (AuNPs) due to an inner filter effect. Aptamers can adsorb on AuNPs to prevent the salt-induced aggregation. AuNPs serve a dual function as fluorescence quencher and colorimetric reporter.
This review (with 110 refs.) gives an overview on the progress that has been made in the past few years on the use of gold nanoparticles (AuNPs) for use in sensors and analytical tools for the determination of dopamine (DA). Both AuNPs and their composites with other organic and inorganic materials including noble metals are treated. Following an overview on the clinical significance of DA, we discuss the various analytical methods that are (a) electrochemiluminescence (ECL); (b) surface enhanced Raman scattering (SERS); (c) colorimetric probing and visual detection; and (d) the large class of electrochemical sensors. Subsections cover sensors based on plain AuNPs, bimetallic NPs, AuNP-metal@metal oxide nanocomposites, AuNP nanocomposites with organic polymers, AuNP nanocomposites with carbon nanotubes or with graphene, and finally sensors based on ternary materials containing AuNPs. The review ends with a conclusion on current challenges of sensors for DA and an outlook on future trends.
We review the recent progress in sensing dopamine based on AuNPs and its nanocomposites including bimetallic nanoparticles, AuNPs-/metal oxide, AuNPs-polymer, AuNPs-carbon nanotubes, AuNPs-graphene and ternary materials using different types of sensing techniques such as electrochemiluminescence (ECL), colorimetric, surface enhanced Raman scattering (SERS) and electrochemical techniques.
The authors describe an electrochemiluminescent (ECL) DNA biosensor that is based on the use of gold nanoparticles (AuNPs) modified with graphite-like carbon nitride nanosheets (g-C3N4 NSs) and carrying a DNA probe. In parallel, nanoparticles prepared from gold-platinum (Au/Pt) alloy and carbon nanotubes (CNTs) were placed on a glassy carbon electrode (GCE). Once the g-C3N4 NHs hybridize with DNA-modified AuNPs, they exhibit strong and stable cathodic ECL activity. The Au/Pt-CNTs were prepared by electrochemical deposition of Au/Pt on the surface of the CNTs in order to warrant good electrical conductivity. On hybridization of immobilized capture probe (S1), target DNA (S2) and labeled signal probe (S3), a sandwich-type DNA complex is formed that produces a stable ECL emission at a typical applied voltage of ?1.18 V and in the presence of peroxodisulfate. Under optimized conditions, the method has a response to target DNA that is linearly related to the logarithm of its concentration in the range between 0.04 f. and 50 pM, with a 0.018 f. detection limit.
The authors introduce a method for spatially arranged DNA immobilization on 10-nm gold nanoparticles (GNP) deposited on a silicon substrate carrying nanogapped interdigitated electrodes. The GNPs are covalently bound to the surface via silane chemistry, and the single steps of fabrication are monitored by FTIR spectroscopy and atomic force microscopy. This GNP deposition technique is shown to reduce the size of the nanogaps to 130 nm. FTIR also was used to monitor the immobilization of DNA on the surface of the interdigitated electrodes. This method allows DNA to be immobilized in a uniform and homogenous way. The utility of the method is demonstrated by immobilizing probe DNA on the surface and detecting target DNA specific for the human papilloma virus via fluorescence with a detection limit as low as 1 pM. In our perception, this method for GNP-mediated DNA immobilization enables high-performance sensing of a wide range of target (analyte) DNA.
Graphical abstract Schematic presentation of gold nanoparticle-mediated and spatially resolved deposition of DNA on nano-gapped interdigitated electrodes. The method was applied to the chemiluminescent determination of the human papillomavirus
A composite material obtained by ultrasonication of graphene oxide (GO) and multi-walled carbon nanotubes (MWCNTs) was loaded with manganese dioxide (MnO2), poly(diallyldimethylammonium chloride) and gold nanoparticles (AuNPs), and the resulting multilayer hybrid films were deposited on a glassy carbon electrode (GCE). The microstructure, composition and electrochemical behavior of the composite and the modified GCE were characterized by transmission electron microscopy, Raman spectra, energy-dispersive X-ray spectroscopy, electrochemical impedance spectroscopy and cyclic voltammetry. The electrode induces efficient electrocatalytic oxidation of dopamine at a rather low working voltage of 0.22 V (vs. SCE) at neutral pH values. The response is linear in the 0.5 μM to 2.5 mM concentration range, the sensitivity is 233.4 μA·mM￣1·cm￣2, and the detection limit is 0.17 μM at an SNR of 3. The sensor is well reproducible and stable. It displays high selectivity over ascorbic acid, uric acid and glucose even if these are present in comparable concentrations.
Graphical abstract Gold nanoparticles were self-assembled onto the surface of the MnO2 decorated graphene oxide-carbon nanotubes composites with poly(diallyldimethylammonium chloride) (PDDA) as a coupling agent. Further, a sensitive electrochemical sensor of dopamine was developed via immobilizing this nanocomposite on a glassy carbon electrode (GCE).
The authors describe an oligonucleotide-based lateral flow test for visual detection of Ag(I). The assay is based on cytosine-Ag(I)-cytosine [C-Ag(I)-C] coordination chemistry to capture gold nanoparticle (AuNP) tags in the test zone. A thiolated C-rich oligonucleotide probe was immobilized on the AuNPs via gold-thiol chemistry, and a biotinylated C-rich oligonucleotide probe was immobilized on the test zone. The AuNPs labelled with C-rich oligonucleotides are captured by Ag(I) ions in the test zone through the C-Ag(I)-C coordination. The resulting accumulation of AuNPs produces a readily visible red band in the test zone. Under optimized conditions, the test is capable of visually detecting 1.0 ppb of Ag(I) which is 50 times lower than the maximum allowable concentration as defined by the US Environmental Protection Agency for drinking water. Hence, the test is inexpensive and highly sensitive. It was applied to the detection of Ag(I) in spiked samples of tap water and river water. In our perception, the test is a particularly valuable tool in limited resource settings.
A method is described for the determination of the pesticide chlorothalonil (CLT). It is based on the finding that citrate-capped gold nanoparticles (AuNPs) undergo aggregation on exposure to chlorothalonil. This is accompanied by a visually detectable color change from wine red to blue. The effect is due to the interaction of the cyano group of chlorothalonil with gold nanoparticles. The assay may also be performed by using a spectrometer. The ratio of absorbances at 700 nm and 520 nm (A700/A520) linearly drops in the 5 to 100 ng·mL?1 CLT concentration range, with a 3.6 ng·mL?1 detection limit. This is below the Chinese guideline value for cucumber. The method is rather simple and does not require any modification of the AuNPs or the utilization of antibody. It was successfully applied to the determination of CLT in (spiked) cucumber samples. Recoveries ranged from 80.4 to 97.4%, and the analytical results compared well with those obtained by HPLC.
Graphical abstract Schematic of the assay. The strong interaction of the cyano group of acetamiprid with gold nanoparticles (AuNPs) via Au-N bond induces the aggregation of gold nanoparticles, and this is accompanied by a color change from red to purple.
The authors describe an electrochemical aptamer based assay for the determination of the serine protease lysozyme in very low (pM) concentrations. The method is based on the formation of a complex between anti-lysozyme aptamer fragments and lysozyme, and on electrochemical detection by differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). The surface of a glassy carbon electrode was modified with a nanocomposite consisting of gold nanoparticles and electrochemically reduced graphene oxide nanosheets (AuNPs/erGO), and the thiolated aptamer was then linked to the AuNPs by self-assembly through Au-S bonds. The interaction of immobilized aptamers with lysozyme leads to the decreased peak current in DPV and increased charge transfer resistance (Rct) in EIS when using hexacyanoferrate or Methylene Blue as a redox probe. The calibration plot, when applying EIS and working at a typical voltage of ?0.22 V (vs. SCE), is linear over 1.0 to 104.3 pM concentration range, with a detection limit of 0.06 pM (at a signal-to-noise ratio of 3). The respective data for DPV are a 9.6–205.5 pM linear range with a detection limit of 0.24 pM. Depending on the redox marker applied, the method works in the “signal-off” or “signal-on” mode in DPV and EIS protocols, respectively. The sensing interface is high specific for lysozyme and not affected by other proteins. The method was applied to the determination of lysozyme in spiked diluted human serum, and the results agreed well with data obtained with a standard ELISA.
Graphical abstract The surface of a glassy carbon electrode was modified with a nanocomposite consisting of gold nanoparticles and electrochemically reduced graphene oxide nanosheets (AuNPs/erGO). Then, the thiolated aptamer was linked to the AuNPs by self-assembly through Au-S bonds. The modified electrode was applied to the determination of lysozyme with “signal off” and “signal on” strategies.
The negatively charged ruthenate(II) complex [Ru(bpy)(PPh3)(CN)3]? and gold nanoparticles (AuNPs) were used for detecting lysozyme (LYS). The luminescence of the ruthenate(II) complex is quenched by AuNPs, and this induces the aggregation of AuNPs and a color change from red to blue. After addition of lysozyme, the positively charged lysozyme and the negatively charged ruthenate(II) complex bind each other by electrostatic interaction firstly. This prevents AuNPs from aggregation and quenches the emission of the ruthenate(II) complex. Its luminescence and the degree of aggregation of the AuNPs can be used to quantify LYS. The fluorometric calibration plot is linear in the 0.01 to 0.20 μM LYS concentration range, and the calibration plot is linear between 0.02 and 0.20 μM of LYS. The color of the solution can be easily distinguished by bare eyes at 0.08 μM or higher concentration of LYS. The applicability of the method was verified by the correct analysis of LYS in chicken egg white.
Graphical abstract Schematic of a luminometric and colorimetric probe based on the induced aggregation of gold nanoparticles by an anionic luminescent ruthenate(II) complex or sensitive lysozyme detection.
An aptamer based assay is described for the colorimetric detection of adenosine. The presence of adenosine triggers the deformation of hairpin DNA oligonucleotide (HP1) containing adenosine aptamer and then hybridizes another unlabeled hairpin DNA oligonucleotide (HP2). This leads to the formation of a double strand with a blunt 3′ terminal. After exonuclease III (Exo III)-assisted degradation, the guanine-rich strand (GRS) is released from HP2. Hence, the adenosine-HP1 complex is released to the solution where it can hybridize another HP2 and initiate many cycles of the digestion reaction with the assistance of Exo III. This leads to the generation of a large number of GRS strands after multiple cycles. The GRS stabilize the red AuNPs against aggregation in the presence of potassium ions. If, however, GRS forms a G-quadruplex, it loses its ability to protect gold nanoparticles (AuNPs) from salt-induced AuNP aggregation. Therefore, the color of the solution changes from red to blue which can be visually observed. This colorimetric assay has a 0.13 nM detection limit and a wide linear range that extends from 5 nM to 1 μM.
Graphical abstract Schematic presentation of a colorimetric aptamer biosensor for adenosine detection based on DNA cycling amplification and salt-induced aggregation of gold nanoparticles.
