The Electrochemiluminescent Immunosensors for Point-of-Care Testing of Methamphetamine Using a Portable Meter

The crowd of methamphetamine (METH) abuse has obviously increased during past decades, their physical and mental health are seriously jeopardized. It compelled growing worry worldwide so that there is an anxious requirement to develop the accurate, sensitive, rapid, and convenient detection techniques to tackle the problem. In regard to this matter, we reported two sensors for METH with synergistical alliance of sensitive electrochemiluminescence (ECL) quantification and specifically immune recognition. These sensors are applicable for point-of-care testing when equipped a portable ECL meter with outstanding performance, makes the feasibility of on-spot analysis. The dual-way analysis of immunosensors can be carried on, both exhibit unprecedented sensitivity toward METH. Here, AuNPs functioned indium tin oxide (ITO) coated polyethylene terephthalate (PET) is used as the basal electrode, providing excellent ECL background and supporting the sensing matrix. The real sample identification demonstrated the practicability and accuracy which met the requirement of forensic judgement and clinical treatment. This work provides a viable methodology and technology for a rapid and reliable analysis of hard drug abuser's samples.     

用于检测癌胚抗原CEA的丝网印刷免疫传感器

采用溶胶-凝胶技术将电子媒介体亚甲基蓝和辣根过氧化物酶(HRP)标记的癌胚抗原(CEA)固定在一次性丝网印刷碳电极表面,制备了CEA免疫传感器。该免疫传感器在含CEA样品的溶液中培育后,抗原-抗体免疫结合物的形成会阻碍HRP活性中心与亚甲基蓝之间的电子传递,使HRP对H2O2电催化氧化的效率降低。循环伏安和计时电流法用于研究免疫电极的电化学特性,在优化的条件下催化效率的降低与CEA质量浓度分别在1.0~6.0μg/L和6.0~138μg/L范围内成线性关系,检出限为0.4μg/L,测定的组内和组间相对标准偏差分别为7.4%和11.2%。该测定无须分离、洗涤步骤,分析时间短,操作方便,检测成本低,具有实际应用价值。     

CdSe Quantum Dots Based Electrochemiluminescence Immunosensor with Simple Structure for Ultrasensitive Detection of Salbutamol

A rapid and ultrasensitive electrochemiluminescence (ECL) competitive immunoassay based on CdSe quantum dots (QDs) and the shorter chain as possible (cysteamine and glutaraldehyde) has been designed for the detection of salbutamol (SAL). Cysteamine and glutaraldehyde made coating antigen immobilize well on the gold electrode surface through the reaction between functional groups, which brought about the simplicity of the immunosensor to some extent. Transmission electron microscopy image, dynamic light scattering, photoluminescence, ultraviolet‐visible absorption and electrochemical impedance spectra were used to characterize the prepared CdSe QDs and the cysteamine/glutaraldehyde/Ovalbumin‐SAL/anti‐SAL‐QDs immunosensor. In the air‐saturated PBS buffer containing 0.1 M K₂S₂O₈ and 0.1 M KCl (pH 9.0), a strong ECL emission of QDs can be observed which depended linearly on the logarithm of the salbutamol concentration with a wide range from 0.05 ng mL^?1 to 100 ng mL^?1, and a detection limit of 0.0056 ng mL^?1. The sensitivity, repeatability, and specificity of the ECL immunosensor have been evaluated. The sensor has been applied to real samples with satisfactory results. This work will open new ways of detecting food additive residue based on QDs ECL in immunoassays.     

Label-free Photoelectrochemical Aptasensor for the Determination of Carcinoembryonic Antigen Using a Cadmum Sulfide Quantum Dot Sensitized Titanium (IV) Oxide Nanotube Electrode

A label-free photoelectrochemical aptasensor for the sensitive and selective determination of carcinoembryonic antigen was constructed based on a CdS quantum dot sensitized TiO₂ nanotube electrode. TiO₂ nanotubes with highly ordered structure and more active sites than bulk TiO₂ were prepared with an electrochemical anodic oxidation process. The CdS quantum dots were immobilized on the TiO₂ nanotubes using poly(diallyldimethylammonium chloride) as a bridge. Due to the energy level match between TiO₂ and CdS, the CdS quantum dots/TiO₂ nanotubes electrode exhibits excellent photoelectrochemical performance. The large surface area of the electrode also allows for capturing large numbers of aptamers. The fine photoelectrochemical performance and the large surface area of the electrode greatly enhanced the detection sensitivity. Under the optimal conditions, the prepared photoelectrochemical aptasensor presents desirable analytical properties for the determination of carcinoembryonic antigen in the range of 0.05 to 10?ng?mL^?1 with a detection limit of 0.014?ng?mL^?1. The application of the designed protocol was investigated by analyzing carcinoembryonic antigen in human serum samples with recoveries from 80.0 to 115.0%. This simple and sensitive method provides an alternative tool to standard biochemical assays.     

Determination of Carcinoembryonic Antigen by Surface-Enhanced Raman Spectroscopy Using Gold Nanobowl Arrays

Surface-enhanced Raman scattering (SERS) based on the double-antibody sandwich format is reported for the determination of carcinoembryonic antigen. Ordered gold nanobowl arrays were fabricated and conjugated with anticarcinoembryonic as capturing substrates, and gold nanoshells, adsorbed with 4-mercaptobenzonic acid, were modified with anticarcinoembryonic antigen as labeling tags. After the carcinoembryonic antigen was captured on ordered gold nanobowl arrays, the labeling tags were bonded to the captured carcinoembryonic antigen. The interaction of SERS substrates (ordered gold nanobowl arrays) and SERS labels (gold nanoshells) showed high sensitivity and a low detection limit for carcinoembryonic antigen. The linear dynamic range of SERS for carcinoembryonic antigen was from 5?pg/mL to 100?ng/mL with a linear relationship between carcinoembryonic antigen concentration and SERS intensity. The detection limit was 1.73?pg/mL. SERS detection may be used for other cancer biomarkers and provides potential for the clinical diagnosis of cancer biomarkers.     

A Facile Electrochemical Immunosensor with Mesoporous Alumina for Detection of Carcinoembryonic Antigen

A label‐free electrochemical immunosensor for the sensitive determination of carcinoembryonic antigen (CEA) was fabricated by immobilizing anti‐CEA onto mesoporous alumina (meso‐Al₂O₃) dispersed in chitosan (0.5 %wt) by the cross‐linking method using glutaraldehyde. Due to its plenty of active sites, meso‐Al₂O₃ showed high catalysis towards hydroquinone. With the electrocatalytic ability of meso‐Al₂O₃ for the reduction of hydroquinone, the current signal of the antigen‐antibody reaction was amplified and the enhanced sensitivity was achieved. The current decreased linearly with CEA concentration in the range of 0.04 to 10 ng/mL (26 pg/mL, S/N=3). The immunosensor had good selectivity and wonderful stability. Furthermore it was applied to the analysis of CEA in serum sample with satisfactory results.     

化学发光免疫分析检测人血清中的癌胚抗原

基于辣根过氧化物酶(HRP)催化H2O2氧化3-(4-羟苯基)丙酸(PHPPA),生成能发荧光的3-(4-羟苯基)丙酸二聚体.在乙腈介质中,在增强剂眯唑参与下,与双[2,4,6-三氯苯基]草酸酯(TCPO)和H2O2反应产生强化学发光.用辣根过氧化物酶(HRP)标记癌胚抗原(CEA)单克隆抗体,通过CEA的双抗夹心免疫...     

A Novel Label-free Chronoamperometric Immunosensor Based on a Biocomposite Material for Rapid Detection of Carcinoembryonic Antigen

In this research, poly(diallyldimethylammonium chloride)-capped gold nanoparticles, nickel ferrite particles, and carbon nanotubes were combined to form a PANC metal composite. The prepared metal composite modified onto a glassy carbon electrode was electropolymerized with poly(o-phenylenediamine) and immobilized with horseradish peroxidase, anti-carcinoembryonic antigen antibody, and bovine serum albumin to create the label-free immunosensors for rapid detection of carcinoembryonic antigen (CEA) using chronoamperometry. This developed biocomposite material modified onto a glassy carbon electrode presented an excellent electrocatalytic response to the redox reaction of hydrogen peroxide as a sensing probe, from which the kinetic parameters including of a charge transfer rate constant, a diffusion coefficient value, an electroactive surface area, and a surface concentration were calculated to be 1.85 s⁻¹, 4.28×10⁻⁶ cm² s⁻¹, 0.14 cm² and 1.87×10⁻⁸ mol cm⁻², respectively. The developed immunosensors also exhibited a wide linear range of CEA concentration from 0.01 to 25 ng mL⁻¹ with high sensitivity (96.21 μA cm⁻² ng⁻¹ mL) and low detection limit (0.72 pg mL⁻¹), excellent selectivity without interfering effects from possible species (amoxicillin, ascorbic acid, aspirin, caffeine, cholesterol, dopamine, glucose, and uric acid), outstanding stability (n=100, %I>50 %), repeatability (%RSD=0.34, n=10), reproducibility (%RSD=4.06, n=10), and rapid analysis (25 s each operation time). This proposed method was successfully applied for CEA detection in whole blood samples with satisfactory results, suggesting that this developed sensing platform may be considered to be exploited for fabrication of other label-free electrochemical immunosensors for the real sample analysis.     

基于碳纳米管修饰电极的胆碱电化学发光生物传感器研制

在碳纳米管(CNTs)和K3Fe(CN)6修饰的铂电极上吸附固定胆碱氧化酶,以鲁米诺为发光试剂,研制了胆碱电化学发光(ECL)生物传感器。CNTs可有效提高电极表面的电荷传输能力、提高电极表面的生物相容性和对酶分子的固载能力;K3Fe(CN)6对酶活性具有激活作用,同时对H2O2增敏的鲁米诺ECL有增强作用,均有利于提高传感器的检测灵敏度。研究表明,将CNTs分散液与K3Fe(CN)6混合,滴涂修饰在Pt电极上,吸附固定胆碱氧化酶,制备传感器。此传感器在含有8×10-6mol/L鲁米诺的磷酸盐缓冲液(pH7.4)、30℃条件下产生的ECL强度与胆碱浓度在1×10-7～4×10-3mol/L范围内呈线性关系,相关系数为0.994,检出限为1.2×10-8 mol/L。此生物传感器应用于鼠血样中胆碱的测定,测得结果为2.68 mg/L,平均回收率为101.1%。传感器具有快速、稳定和重现性好等特点,有望应用于常规分析。     

Highly sensitive luminol electrochemiluminescence immunosensor based on ZnO nanoparticles and glucose oxidase decorated graphene for cancer biomarker detection

In this work, we reported a sandwiched luminol electrochemiluminescence (ECL) immunosensor using ZnO nanoparticles (ZnONPs) and glucose oxidase (GOD) decorated graphene as labels and in situ generated hydrogen peroxide as coreactant. In order to construct the base of the immunosensor, a hybrid architecture of Au nanoparticles and graphene by reduction of HAuCl₄ and graphene oxide (GO) with ascorbic acid was prepared. The resulted hybrid architecture modified electrode provided an excellent platform for immobilization of antibody with good bioactivity and stability. Then, ZnONPs and GOD functionalized graphene labeled secondary antibody was designed for fabricating a novel sandwiched ECL immunosensor. Enhanced sensitivity was obtained by in situ generating hydrogen peroxide with glucose oxidase and the catalysis of ZnONPs to the ECL reaction of luminol–H₂O₂ system. The as-prepared ECL immunosensor exhibited excellent analytical property for the detection of carcinoembryonic antigen (CEA) in the range from 10 pg mL⁻¹ to 80 ng mL⁻¹ and with a detection limit of 3.3 pg mL⁻¹ (S N⁻¹ = 3). The amplification strategy performed good promise for clinical application of screening of cancer biomarkers.     

基于碳纳米管修饰电极的胆碱电化学发光生物传感器研制

在碳纳米管(CNTs)和K3Fe(CN)6修饰的铂电极上吸附固定胆碱氧化酶,以鲁米诺为发光试剂,研制了胆碱电化学发光(ECL)生物传感器.CNTs可有效提高电极表面的电荷传输能力、提高电极表面的生物相容性和对酶分子的固载能力;K3Fe(CN)6对酶活性具有激活作用,同时对H2O2增敏的鲁米诺ECL有增强作用,均有利于提...     

A Novel Electrochemical Aptasensor for Carcinoembryonic Antigen Detection Based on Target‐induced Bridge Assembly

The present study describes a novel electrochemical aptasensor for detection of carcinoembryonic antigen (CEA), a key cancer biomarker. The sensing strategy relied on the CEA‐induced bridge assembly, as a physical barrier, on the surface of gold electrode, resulting in a significant increase of the sensor sensitivity. Under optimal conditions, the aptasensing platform showed a wide linear range (3 pg/mL to 40 ng/mL) and a low detection limit (0.9 pg/mL). Some possible interfering materials were also assessed and the results indicated that the designed aptasensor had good specificity toward CEA. The quantitation of CEA in the spiked human serum samples confirmed the reliability and applicability of the electrochemical aptasensor. So, the developed sensing method has a potential application in the clinical diagnosis.     

Electroanalytical Determination of Carcinoembryonic Antigen at a Silica Nanoparticles/Titania Sol–Gel Composite Membrane‐Modified Gold Electrode

A highly hydrophilic and nontoxic colloidal silica nanoparticle/titania sol–gel composite membrane was prepared on a gold electrode via a chemical vapor deposition method. With carcinoembryonic antigen (CEA) as a model antigen and encapsulation of carcinoembryonic antibody (anti‐CEA) in the composite architecture, this membrane could be used for reagentless electrochemical immunoassay. The presence of silica nanoparticles provided a congenial microenvironment for adsorbed biomolecules. The formation of immunoconjugate by a simple one‐step immunoreaction between CEA in sample solution and the immobilized anti‐CEA introduced the change in the potential. The modified procedure was further characterized by electrochemical impedance spectroscopy and cyclic voltammetry. Compared to the commonly applied methods, i.e., the TiO₂ direct embedding procedure, this strategy could allow for antibodies immobilized with higher loading amount and better retained immunoactivity. The resulting immunosensor exhibited high sensitivity, good precision, acceptable stability, accuracy, reproducibility and wide linear range from 1.5 to 240 ng mL^?1 with a detection limit of 0.5 ng mL^?1 at 3σ. Analytical results of clinical samples show that the developed immunoassay is comparable with the enzyme‐linked immunosorbent assays (ELISAs) method, implying a promising alternative approach for detecting CEA in the clinical diagnosis. Furthermore, this composite membrane could be used efficiently for the entrapment of other biomarkers and clinical applications.     

水溶性量子点荧光探针用于胃癌细胞相关抗原CA242的检测

基于量子点荧光探针对胃癌细胞相关抗原CA242进行了检测。首先在水溶液中直接合成性能优良的量子点荧光纳米颗粒,并在其表面成功修饰了羊抗小鼠IgG和聚乙二醇,制得功能化的水溶性量子点荧光探针,并利用探针对胃癌细胞相关抗原CA242进行检测,进一步与传统的基于荧光染料标记的免疫荧光分析方法进行了比较。实验结果表明:该功能化的探针能够有效地识别胃癌细胞相关抗原CA242,并且在光稳定性和灵敏度方面都较传统的基于荧光染料标记的免疫荧光分析方法有明显的改善,从而为CA242的相关检测以及胃癌的诊断与愈后判断提供了新的方法。     

Construction, Expression and Characterization of a Chimeric Protein Targeting Carcinoembryonic Antigen in Lung Cancer

The carcinoembryonic antigen(CEA) is an oncofetal glycoprotein known as an important clinical tumor marker and is overexpressed in several types of tumors, including colorectal and lung carcinomas. We constructed a chimeric protein that exhibits both specific binding and immune stimulating activities, by fusing staphylococcal enterotoxin A(SEA) to the C-terminus of an anti-CEA single-chain disulfide-stabilized Fv(scdsFv) antibody (single-chain-C-terminus/SEA, SC-C/SEA). The SC-C/SEA protein was expressed in...     

A Novel Strategy for Simultaneous Determination of Dopamine and Uric Acid Using a Carbon Paste Electrode Modified with CdTe Quantum Dots

A novel CdTe quantum dots‐modified carbon paste electrode (QDMCPE) was fabricated and used to study the electrooxidation of dopamine and uric acid and their mixtures by electrochemical methods. Using square wave voltammetry (SWV), a highly sensitive and simultaneous determination of dopamine and uric acid was explored at the modified electrode. SWV peak currents of dopamine and uric acid increased linearly with their concentrations in the ranges of 7.5×10^?8–6.0×10^?4 M, and 7.5×10^?6–1.4×10^?3 M, respectively. Finally this new sensor was used for determination of dopamine and uric acid in some real samples.     

Electrochemiluminescence (ECL) Detection of Ammonium Ion Based on a Novel Iridium Complex Modified Electrode

We report that ammonium ion, as its organic amine counterparts, can be sensitively detected as a coreactant in the electrochemiluminescence (ECL) reaction of a novel iridium complex (pq)₂Ir(N-phMA), where pq is 2-phenylquinoline anion and N-phMA is N-phenylmethacrylamide. The modified ECL electrode was fabricated by casting deposition of a homogeneous MWNTs/PVA/(pq)₂Ir(N-phMA) dispersion solution onto the surface of a glassy carbon electrode. The electrode responds sensitively to ammonium ion in a wide concentration range. Two regression equations were established: Y = 0.7016X + 46.8 (R² = 0.9985) and Y = 0.2565X + 174.2 (R² = 0.9991) for the concentration from 1–180 nM and 180–1800 nM, respectively. The limit of detection was as low as 0.13 nM (S/N = 3).     

Application of ZnO quantum dots dotted carbon nanotube for sensitive electrochemiluminescence immunoassay based on simply electrochemical reduced Pt/Au alloy and a disposable device

We report on a disposable microdevice suitable for sandwich-type electrochemiluminescence (ECL) detection of prostate specific antigen (PSA). The method is making use of ZnO quantum dots dotted carbon nanotube (ZnO@CNT) and simply electrochemical reduced Pt/Au alloy. The latter was selected as immunosensing probe to modify screen-printed carbon electrode, due to its excellent electrical property. For further ultrasensitive, low-potential and stable ECL detection, ZnO@CNT composite was first synthesized using a facile solvothermal method, and employed as signal amplification label. In this work, two working electrodes in one device were used for one determination to obtain more exact results based on screen-print technique. Taking advantage of dual-amplification effects of the Pt/Au and ZnO@CNT, this immunosensor could detect the PSA quantitatively, in the range of 0.001–500 ng mL⁻¹, with a low detection limit of 0.61 pg mL⁻¹. The resulting versatile immunosensor possesses high sensitivity, satisfactory reproducibility and regeneration. This simple and specific strategy has vast potential to be used in other biological assays.     

Nitrogen-doped TiO2 Nanocrystals for Highly Sensitive Electrochemical Immunoassay of Carcinoembryonic Antigen

Nitrogen-doped TiO₂ nanocrystals (N−TiO₂ NCs) were simply synthesized and then functionalized with streptavidin for highly sensitive electrochemical immunoassay of tumor marker. Scanning electron microscopy, transmission electron microscopy, static water contact angle, and cyclic voltametric measurement were adapted to examine the properties of N−TiO₂ NCs and resultant immunosensor. The functionalized N−TiO₂ NCs sensing platform shows high electrochemical conductivity, large surface area and excellent hydrophilicity. The features make them to produce high current response, capture more antibody molecules, and maintain the bioactivity of immobilized antibodies. By means of carcinoembryonic antigen (CEA) as model tumor marker, a wide linear range of 0.005–3 ng/mL and a low detection limit of 0.005 ng/mL (signal-to-noise ratio of 3) were achieved by the proposed CEA immunosensor. Furthermore, the resultant CEA immunosensor displays high specificity and was employed to determine CEA in clinical serum samples.     

电化学发光免疫传感器检测甲磺隆的研究

电化学发光 ( Electrochemiluminescence,ECL)分析是对电极施加一定的电压进行电化学反应 ,反应的产物之间或与体系中某种组分发生化学发光 ,用普通光学手段测量发光光谱和强度从而对物质进行痕量分析的一种方法 .与其它检测方法相比 ,它具有一些明显的优势 :标记物的检出限低 ( 2 0 0fmol/L ) ;动力学范围宽达 6个数量级 ;标记物比大多数化学发光标记物稳定 ;由于是电促发光 ,只有靠近电极表面的带有标记物的部分才能被检测到 ,所以分离或非分离体系均可应用此方法 .电化学发光常用的标记试剂分子是联吡啶钌 [Ru( bpy) 2 + 3].Ru( bpy) …