Electrochemiluminescence Aptasensor for the MUC1 Protein Based on Multi‐functionalized Graphene Oxide Nanocomposite

The electrochemiluminescence (ECL) aptasensor was prepared for the detection of Mucin 1 based on its specific recognition by aptamer immobilized on multi‐functionalized graphene oxide nanocomposite, which was prepared with N‐(4‐aminobutyl)‐N‐ethylisoluminol (ABEI) and aptamer chemically bound to the surface of magnetic GO (nanoFe₃O₄@GO). ABEI and aptamer acted as the electrochemiluminophore and the capture device for Mucin 1 respectively. NanoFe₃O₄@GO brought multi‐functionalized graphene oxide nanocomposite attracted on the surface of magnetic glass carbon electrode through magnetism, enabled all the ABEI immobilized electrochemically active due to its good conductivity and thus then facilitated the sensitive detection of Mucin 1. In addition, the ECL aptasensor can be prepared through a one‐step process. Under optimal conditions, the ECL intensity of the aptasensor decreased proportionally to the logarithmic concentrations of Mucin 1 in the range of 0.005–1000 ng mL^?1. This aptasensor displays good specificity, stability, reproducibility and application. This method has a large potential because such a multi‐functionalized graphene oxide nanocomposite also may be applied to other ECL‐based aptasensors.     

Iodide Recognition and Sensing in Water by a Halogen‐Bonding Ruthenium(II)‐Based Rotaxane

The synthesis and anion‐recognition properties of the first halogen‐bonding rotaxane host to sense anions in water is described. The rotaxane features a halogen‐bonding axle component, which is stoppered with water‐solubilizing permethylated β‐cyclodextrin motifs, and a luminescent tris(bipyridine)ruthenium(II)‐based macrocycle component. ¹H NMR anion‐binding titrations in D₂O reveal the halogen‐bonding rotaxane to bind iodide with high affinity and with selectively over the smaller halide anions and sulfate. The binding affinity trend was explained through molecular dynamics simulations and free‐energy calculations. Photo‐physical investigations demonstrate the ability of the interlocked halogen‐bonding host to sense iodide in water, through enhancement of the macrocycle component’s Ru^II metal–ligand charge transfer (MLCT) emission.     

Electrogenerated Chemiluminescence Sensor Based on Tris(2,2′‐bipyridine)ruthenium(II)‐Immobilized Natural Clay and Ionic Liquid

A novel electrogenerated chemiluminescence (ECL) sensor based on natural clay and ionic liquid was fabricated. Tris(2,2′‐bipyridine)ruthenium(II) (Ru(bpy)₃²⁺) was immobilized on natural clay surface through simple adsorption. An ECL sensor was prepared by mixing Ru(bpy)₃²⁺‐incorporated clay, graphite powder and an ionic liquid (1‐butyl‐3‐methylimidazolium hexafluorophosphate) as the binder. The electrochemical behavior and ECL of the immobilized Ru(bpy)₃²⁺ was investigated. It was observed that the ECL of immobilized Ru(bpy)₃²⁺ was activated by the ionic liquid. The proposed ECL sensor showed high sensitivity to tri‐n‐propylamine (TPrA) and the detection limit was found to be 20 pM. In addition, the ECL sensor displayed good stability for TPrA detection and long‐term storage stability.     

A Bifunctional Electrocatalyst Containing Tris(2,2′‐bipyridine) Ruthenium(II) and 12‐Molybdophosphate Bulk‐Modified Carbon Paste Electrode

The electroactive composite containing tris(2,2′‐bipyridine) ruthenium(II) and 12‐molybdophosphate (RuPMo₁₂) was synthesized and first used as a bifunctional electrocatalyst to fabricate a chemically bulk‐modified carbon paste electrode (RuPMo₁₂‐CPE) by direct mixing. The electrochemical behavior of the RuPMo₁₂‐CPE was studied by cyclic voltammetry. The RuPMo₁₂‐CPE presents good electrocatalytic activity not only toward the reduction of hydrogen peroxide and bromate, which is attributed to the function of molybdophosphate, but also toward the oxidation of arsenite, which is primarily attributed to the function of tris(2,2′‐bipyridine) ruthenium(II). The remarkable advantage of the RuPMo₁₂‐CPE is its good stability owing to the insolubility of RuPMo₁₂ and reproducibility of surface renewal.     

Label‐ and modification‐free‐based in situ selection of bovine serum albumin specific aptamer

Systematic evolution of ligands by exponential enrichment is a traditional approach to select aptamer, which has a great potential in biosensing field. However, chemical modifications of DNA library or targets before selection might block the real recognition and binding sites between aptamers and their targets. In this study, a label‐ and modification‐free‐based in situ selection strategy was developed to overcome this limitation. The strategy is an attempt to screen bovine serum albumin aptamers according to the principle of electrophoretic mobility shift assay, and allowed single‐stranded DNA sequence to be fully exposed to interact with bovine serum albumin which was mixed with the agarose gel beforehand. After eight rounds of selection, specific aptamer with low dissociation constant (K_d) value of 69.44 ± 7.60 nM was selected and used for subsequent establishment of fluorescence biosensor. After optimization, the optimal aptasensor exhibited a high sensitivity toward bovine serum albumin with a limit of detection of 0.24 ng/mL (linear range from 1 to 120 ng/mL). These results indicated that the label‐ and modification‐free‐based in situ selection strategy proposed in this work could effectively select specific aptamer to develop aptasensor for sensitive detection of bovine serum albumin or other targets in actual complicated samples.     

Layer‐by‐layer Assembled Multilayers of Graphene/Mono‐(6‐amino‐6‐deoxy)‐β‐cyclodextrin for Detection of Dopamine

Graphene/mono‐(6‐amino‐6‐deoxy)‐β‐cyclodextrin multilayer films composed of graphene sheet (GS) and mono‐(6‐amino‐6‐deoxy)‐β‐cyclodextrin (NH₂‐β‐CD) were fabricated easily by two steps. First, negatively charged graphene oxide (GO) and positively charged mono‐(6‐amino‐6‐deoxy)‐β‐cyclodextrin (NH₂‐β‐CD) were layer‐by‐layer (LBL) self‐assembled on glassy carbon electrode (GCE) modified with a layer of poly(diallyldimethylammonium chloride) (PDDA). Then graphene/mono‐(6‐amino‐6‐deoxy)‐β‐cyclodextrin (GS/NH₂‐β‐CD) multilayer films were built up by electrochemical reduction of graphene oxide/mono‐(6‐amino‐6‐deoxy)‐β‐cyclodextrin (GO/NH₂‐β‐CD). Combining the high surface area of GS and the active recognition sites on β‐cyclodextrin (β‐CD), the GS/NH₂‐β‐CD multilayer films show excellent electrochemical sensing performance for the detection of DA with an extraordinary broad linear range from 2.53 to 980.05 µmol·L^?1. This study offers a simple route to the controllable formation of graphene‐based electrochemical sensor for the detection of DA.     

A Sandwich‐type Electrochemiluminescence Aptasensor for Thrombin Based on Functional Co‐polymer Electrode Using Ru(bpy)32+ Doped Nanocomposites as Signal‐amplifying Tags

Herein, a signal‐on sandwich‐type electrochemiluminescence (ECL) aptasensor for the detection of thrombin (TB) was proposed. The graphene (GR) doped thionine (TH) was electropolymerized synchronously on the bare glassy carbon electrode (GCE) to form co‐polymer (PTG) electrode. The gold nanoparticles (AuNPs) were decorated on the surface of the PTG by in‐situ electrodeposition, and the functional co‐polymer (PTG‐AuNPs) electrode was utilized as sensing interface. Then, TB binding aptamer I (TBA I) as capture probes were modified on the PTG‐AuNPs electrode to capture TB, and Ru(bpy)₃²⁺/silver nanoparticles doped silica core‐shell nanocomposites‐labeled TB binding aptamer II (RuAg/SiO₂NPs@TBA II) were used as signal probes to further bind TB, resulting in a sandwich structure. With the assistant of silica shell and AgNPs, the enrichment and luminous efficiency of Ru(bpy)₃²⁺ were significantly improved. Under the synergy of PTG‐AuNPs and RuAg/SiO₂NPs, the ECL signal was dramatically increased. The proposed ECL aptasensor displayed a wide linear range from 2 fM to 2 pM with the detection limit of 1 fM, which is comparable or better than that in reported ECL aptasensors for TB using Ru(bpy)₃²⁺ and its derivatives as the luminescent substance. The excellent sensitivity makes the proposed aptasensor a promising potential in pharmaceutical and clinical analysis.     

Paper‐Based Electrochemiluminescent 3D Immunodevice for Lab‐on‐Paper,Specific, and Sensitive Point‐of‐Care Testing

Recent research on microfluidic paper‐based analytical devices (μPADs) has shown that paper has great potential for the fabrication of low‐cost diagnostic devices for healthcare and environmental monitoring applications. Herein, electrochemiluminescence (ECL) was introduced for the first time into μPADs that were based on screen‐printed paper‐electrodes. To further perform high‐specificity, high‐performance, and high‐sensitivity ECL on μPADs for point‐of‐care testing (POCT), ECL immunoassay capabilities were introduced into a wax‐patterned 3D paper‐based ECL device, which was characterized by SEM, contact‐angle measurement, and electrochemical impedance spectroscopy. With the aid of a home‐made device‐holder, the ECL reaction was triggered at room temperature. By using a typical tris(bipyridine)ruthenium–tri‐n‐propylamine ECL system, this paper‐based ECL 3D immunodevice was applied to the diagnosis of carcinoembryonic antigens in real clinical serum samples. This contribution further expands the number of sensitive and specific detection modes of μPADs.     

A sensitive, non-damaging electrochemiluminescent aptasensor via a low potential approach at DNA-modified gold electrodes

Electrochemiluminescence (ECL)-based biosensors are often used in the field of DNA- and protein-assay. Although ruthenium complex-based ECL is sensitive, its high exciting potential may lead to oxidation damage to biomolecules. For the first time, a non-damaging, low potential ECL aptasensor was constructed for bioassay with lysozyme as a model. After a single-stranded anti-lysozyme aptamer was attached to a gold electrode, a double stranded (ds)-DNA formed with its complementary strand. Ru(phen)(3)(2+), as an ECL probe, was intercalated into the ds-DNA. The hybridization of lysozyme with its aptamer led to the dissociation of ds-DNA because of the high stability of the aptamer-lysozyme and therefore the Ru(phen)(3)(2+) intercalated into ds-DNA was released. A low potential ECL was observed at the ds-DNA-modified electrode because ds-DNA was able to preconcentrate tripropylamine (TPA) and acted as the acceptor of the protons released from protonated TPAH(+). While the DNA sequence (anti-lysozyme aptamer) was used as the special recognition element for lysozyme, the formed ds-DNA also provided a micro-environment for low potential ECL. The low potential ECL aptasensor achieved the determination of lysozyme with a detection limit of 0.45 pM. The day-to-day precision (RSDs, n = 5) for the determination of lysozyme was lower than 5%, showing the reliability of the aptasensor. The regeneration of the aptasensor confirmed that the low potential for ECL could decrease oxidation damage to biomolecules. Further, the proposed method was successfully used to analyze diluted egg white sample directly. The protocol exhibited a promising platform for sensitive bioassay and could be further applied for the development of other low potential ECL sensing systems.     

基于Ru(bpy)32+-SiO2纳米颗粒的核酸适配体传感器对凝血酶的电致化学发光检测

本文应用核酸适配体构建了一种新型的电致化学发光检测蛋白体系。两个核酸适配体结合凝血酶的两个不同位点,利用这两核酸适配体与凝血酶的高亲和力构建三明治传感体系检测凝血酶。一个核酸适配体固定在金电极上用来捕获凝血酶,另一个标记有包裹电致化学发光活性物Ru(bpy)₃²⁺的二氧化硅纳米颗粒,用来检测电致化学发光信号。此核酸适配体传感器对凝血酶具有特异识别性,电致化学发光信号与凝血酶的浓度直接相关,非特异性识别的牛血红蛋白、牛血清白蛋白不干扰测定。由于在检测的核酸适配体上标记的纳米颗粒包裹有多个发光活性物,因此大大提高了发光效率和灵敏度,此法对凝血酶的线性响应范围为2.0 fmol•L^-1～2.0 pmol•L^-1,检测限可达1.0 fmol•L^-1。     

Mixed iridium(III) and ruthenium(II) polypyridyl complexes containing poly(ε‐caprolactone)‐bipyridine macroligands

A hydroxy‐functionalized bipyridine ligand was polymerized with ε‐caprolactone utilizing the controlled ring‐opening polymerization of ε‐caprolactone in the presence of stannous octoate. The resulting poly(ε‐caprolactone)‐containing bipyridine was characterized by ¹H NMR and IR spectroscopy, and gel permeation chromatography, as well as matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry, revealing the successful incorporation of the bipyridine ligand into the polymer chain. Coordination to iridium(III) and ruthenium(II) precursor complexes yielded two macroligand complexes, which were characterized by NMR, gel permeation chromatography, matrix‐assisted laser desorption/ionization time‐of‐flight MS, cyclic voltammetry, and differential scanning calorimetry. In addition, both photophysical and electrochemical properties of the metal‐containing polymers proved the formation of a trisruthenium(II) and a trisiridium(III) polypyridyl species, respectively. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 4153–4160, 2004     

Self‐assembly of amphiphilic tris(2,2′‐bipyridine)ruthenium‐cored star‐shaped polymers

Amphiphilic tris(2,2′‐bipyridine)ruthenium‐cored star‐shaped polymers consisting of one polystyrene block and two poly(N‐isopropylacrylamide) blocks were prepared by the “arm‐first” method in which RAFT polymerization and nonconvalent ligand–metal complexation were employed. The prepared amphiphilic star‐shaped metallopolymers are able to form micelles in water. The size and distribution of the micelles were studied by dynamic light scattering and transmission electron microscopy techniques. Preliminary studies indicate that the polymer concentration and the hydrophilic poly(N‐isopropylacrylamide) block length can affect the morphologies of the formed metal‐interfaced core–shell micelles in water. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 45: 4204–4210, 2007     

Luminescent polymeric ruthenium complexes with polystyrene‐b‐poly(methyl methacrylate) macroligands: The sequential activation of initiator sites for blocks generated by parallel polymerization mechanisms

The synthesis of polystyrene‐b‐poly(methyl methacrylate) diblock copolymers with a luminescent ruthenium(II) tris(bipyridine) [Ru(bpy)₃] complex at the block junction is described. The macroligand precursor, polystyrene bipyridine‐poly(methyl methacrylate) [bpy(PS–H)(PMMA)], was synthesized via the atom transfer radical polymerization of styrene and methyl methacrylate from two independent, sequentially activated initiating sites. Both polymerization steps resulted in the growth of blocks with sizes consistent with monomer loading and narrow molecular weight distributions (i.e., polydispersity index < 1.3). Subsequent reactions with ruthenium(II) bis(bipyridine) dichloride [Ru(bpy)₂Cl₂] in the presence of Ag⁺ generated the ruthenium tris(bipyridine)‐centered diblock, which is of interest for the imaging of block copolymer microstructures and for incorporation into new photonic materials. © 2002 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 40: 4250–4255, 2002     

Ru catalyzed cyclodimerization of bis(2‐thienyl)acetylene and bis(3‐thienyl)acetylene. Synthesis properties of 4,5,6‐tris(2‐thienyl)benzo(b)thiophene and 5,6,7‐tris(3‐thienyl)benzo[b]thiophene

Activated dihydridocarbonyltris(triphenylphosphine)ruthenium catalyzes the cyclodimerization of both bis(2‐thienyl)acetylene and bis(3‐thienyl)acetylene to yield, respectively, 4,5,6‐tris(2′‐thienyl)‐benzo[b]thiophene and 5,6,7‐tris(3′‐thienyl)benzo[b]thiophene. These fluoresce in the blue. Both undergo irreversible one electron oxidation at & sim1.1 mV versus Ag/Ag⁺ electrode, consistent with oxidation of the benzo[b]thiophene nuclei rather than the substituent thiophene rings.     

Electrochemiluminescence of Tris(2,2′‐bipyridine)ruthenium with Various Co‐reactants under Ultrasound Irradiation

Electrochemiluminescence (ECL) of tris(2,2′‐bipyridine)ruthenium, Ru(bpy)₃²⁺ in the presence of various co‐reactants, such as tripropylamine (TPA), oxalate ion (C₂O₄^2?), ascorbic acid (H₂A) and dehydroascorbic acid (DHA), were investigated under ultrasound irradiation. In sono‐ECL experiments, an indium‐thin‐oxide (ITO) was used as working electrode, and a titanium tipped sonic horn probe (diameter 2 mm) which operated at a frequency of 20 kHz was set in the front of the ITO electrode. Under the ultrasound irradiation, ECL signals were found to be significantly enhanced when TPA and C₂O₄^2? were used as co‐reactants, only slightly enhanced in Ru(bpy)₃²⁺/DHA system, but total quenched in Ru(bpy)₃²⁺/H₂A system. The difference of Ru(bpy)₃²⁺ ECL behaviors for various co‐reactant could to be due to the different kinetics of catalytic reactions associated in ECL schemes. ECL quenching effect observed in Ru(bpy)₃²⁺/H₂A system was suggested to be due to electron transfer (ET) route between the excited state *Ru(bpy)₃²⁺ and ascorbate anion HA^? diffused from the bulk solution, where the diffusional HA^? species served as electron donor. The effect becomes more pronounced upon sonication because the effective collision frequency between *Ru(bpy)₃²⁺ and HA^? would be significantly increased by the enhanced mass transport effect of ultrasound.     

Graphene oxide based molecularly imprinted polymers modified with β‐cyclodextrin for selective extraction of di(2‐ethylhexyl) phthalate in environmental waters

Graphene oxide based molecularly imprinted polymers modified with β‐cyclodextrin were prepared as solid‐phase extraction column sorbents for specific recognition and sensitive detection of di(2‐ethylhexyl) phthalate in water samples. The morphology and composition of synthesized sorbents were characterized by scanning electron microscopy, thermo‐gravimetric analysis, Raman spectroscopy, and Fourier‐transform infrared spectroscopy. The conditions affecting the performance of extraction procedures such as desorption solvent types and volume, sample pH and volume were investigated. The loading capacity (8.2 μg/mg) of the prepared sorbents increased eight times after modification with β‐cyclodextrin. The developed extraction procedures coupled to high‐performance liquid chromatography exhibited good linearity (0.2–500 μg/L), low limit of detection (0.052 μg/L), and good precision (relative standard deviation?5.7%) under optimized conditions. The developed solid‐phase extraction technique with prepared sorbents has been successfully applied in extracting trace di(2‐ethylhexyl) phthalate from real natural waters with high efficiency, good selectivity, and desirable recoveries.     

Detection of thrombin using electrogenerated chemiluminescence based on Ru(bpy)3(2+)-doped silica nanoparticle aptasensor via target protein-induced strand displacement

A sensitive and selective aptasensor using tri(2,2′-bipyridyl)ruthenium(II)-doped silica nanoparticles (Ru(bpy)₃²⁺-doped SNPs) as DNA tags for detection of thrombin is developed based on the target protein-induced strand displacement of the DNA probe. For the proposed aptasensor, the aptamer was assembled on the surface of the Au electrode through Au-S binding. The hybridization event between the DNA probe labeled by the Ru(bpy)₃²⁺-doped SNPs and the aptamer was evaluated by electrogenerated chemiluminescence (ECL) measurements. Then, the DNA probe was displaced by thrombin and the binding event between the thrombin and the aptamer was monitored by ECL measurements again. The difference of ECL intensity (ΔI_ECL) of the two events could be used to quantify the thrombin. Other proteins, such as bovine serum albumin and bovine hemoglobin, had almost negligible ΔI_ECL. Under the optimal conditions, the ΔI_ECL was linearly related to the concentration of the thrombin in the range of 10 fM to 10 pM and the detection limit was down to 1.0 fM since SNPs containing a large number of Ru(bpy)₃²⁺ molecules were labeled on the DNA probe.     

Synthesis and photophysical properties of novel amphiphilic ruthenium (II) complexes containing 4,4′‐dialkylaminomethyl‐2,2′‐bipyridyl ligands

The synthesis of a number of new 2,2′‐bipyridine ligands functionalized with bulky amino side groups is reported. Three homoleptic polypyridyl ruthenium (II) complexes, [Ru(L)₃]²⁺ 2(PF₆^?), where L is 4,4′‐dioctylaminomethyl‐2,2′‐bipyridine (Ru4a), 4,4′‐didodecylaminomethyl‐2,2′‐bipyridine (Ru4b) and 4,4′‐dioctadodecylaminomethyl‐2,2′‐bipyridine (Ru4c), have been synthesized. These compounds were characterized and their photophysical properties examined. The electronic spectra of three complexes show pyridyl π → π* transitions in the UV region and metal‐to‐ligand charge transfer bands in the visible region. Copyright © 2005 John Wiley & Sons, Ltd.     

Tris(2,2′‐bipyridyl)ruthenium(II) Electrogenerated Chemiluminescence Sensor Based on Platinized Carbon Nanotube–Zirconia–Nafion Composite Films

Mesoporous films of platinized carbon nanotube–zirconia–Nafion composite have been used for the immobilization of tris(2,2′‐bipyridyl)ruthenium (II) (Ru(bpy)₃²⁺) on an electrode surface to yield a solid‐state electrogenerated chemiluminescence (ECL) sensor. The composite films of Pt–CNT–zirconia–Nafion exhibit much larger pore diameter (3.55 nm) than that of Nafion (2.82 nm) and thus leading to much larger ECL response for tripropylamine (TPA) because of the fast diffusion of the analyte within the films. Due to the conducting and electrocatalytic features of CNTs and Pt nanoparticles, their incorporation into the zirconia–Nafion composite films resulted in the decreased electron transfer resistance within the films. The present ECL sensor based on the Pt–CNT–zirconia–Nafion gave a linear response (R²=0.999) for TPA concentration from 3.0 nM to 1.0 mM with a remarkable detection limit (S/N=3) of 1.0 nM, which is much lower compared to those obtained with the ECL sensors based on other types of sol‐gel ceramic–Nafion composite films such as silica–Nafion and titania–Nafion.     

Synthesis of N,N,N-4,4＂-Di-（4-methylphenyl）-2,2＇：6＇,2＂- terpyridine-N,N,N-tris（isothiocyanato） Ruthenium（Ⅱ） and Application to Colorimetric Hg^2＋ Sensor

A terpyridine derivative DPTP [di-（4-methylphenyl）-2,2＇：6＇,2＂-terpyridine] was conveniently synthesized from 2-bromopyridine via halogen-dance reaction, Kharash coupling and Stille coupling reaction. Then its corresponding ruthenium complex Ru-DPTP [N,N,N-4,4＇＇-di-（4-methy,phenyl）-2,2＇：6＇,2＂-terpyridine-N,N,N-tris（is,-thi,cyanat,）- ruthenium（H） ammonium] was obtained and fully characterized by IR, UV-Vis, ESI MS and elemental analysis. The MLCT absorption band of Ru-DPTP was blue-shifted from 570 to 500 nm upon addition of Hg^2＋. Among a series of surveyed metal ions, the complex showed a unique recognition to Hg^2＋, indicating that it can be used as a selective colorimetric sensor for Hg^2＋.