多孔硅适配子生物传感器的电化学研究

构建1种用于快速检测四环素的新型电化学纳米多孔硅(PS)生物传感器。通过脉冲腐蚀法制得多孔硅基片,将适配子固定于其上,这种四环素适配子能够特异性识别四环素分子,并引起阻抗值的变化。利用电化学交流阻抗法比较固定适配子前后硅片表面阻抗值的变化,以及在体系中加入不同浓度四环素后阻抗谱的变化。选择1个合适的等效电路对测得的阻抗数据进行拟合,获得了四环素浓度与阻抗值的变化规律。传感器的线性检测范围为2.079~62.37 nmol/L,检测限为2.079 7 nmol/L。     

多孔硅Bragg反射镜适配子生物传感器检测心肌肌钙蛋白Ⅰ

通过脉冲腐蚀法制备多孔硅Bragg反射镜,将心肌肌钙蛋白Ⅰ(cTnⅠ)适配子共价固定到多孔硅Bragg反射镜的孔洞中,发现适配子能与cTnⅠ分子特异性结合.定量分析不同浓度的cTnⅠ与适配子结合后多孔硅Bragg反射镜的反射谱峰位的红移情况.结果表明:基于多孔硅Bragg反射镜适配子生物传感器的光学检测具有良好的特异性,且具有免标记及检测时间短等优异性能.传感器的线性检测范围0.05～4 nmol/L,最低检测限为0.05 nmol/L.     

多孔硅Bragg反射镜适配子生物传感器检测心肌肌钙蛋白I

通过脉冲腐蚀法制备多孔硅Bragg反射镜,将心肌肌钙蛋白I（cTnI）适配子共价固定到多孔硅Bragg反射镜的孔洞中,发现适配子能与cTnI分子特异性结合。定量分析不同浓度的cTnI与适配子结合后多孔硅Bragg反射镜的反射谱峰位的红移情况。结果表明：基于多孔硅Bragg反射镜适配子生物传感器的光学检测具有良好的特异性,且具有免标记及检测时间短等优异性能。传感器的线性检测范围0．05-4nmol／L,最低检测限为0．05nmol／L。     

适配子纳米生物传感器在青霉素检测中的应用

通过脉冲腐蚀法对硅片进行多孔硅的制备,利用玻片通过对共价法、离子吸附法和APTES修饰的戊二醛交联法3种固定适配子方法的对比,以确定较好的固定青霉素适配子的方法。将适配子固定在多孔硅上后,利用交流阻抗法对加入青霉素前后传感器阻抗值进行测定、对比,构建等效电路并进行阻抗拟合。对多孔硅传感器的Nyqu ist谱图进行分析以确定多孔硅表面成功固定了青霉素适配子,从而证明构建纳米生物传感器成功。传感器的线性检测范围为0.05~0.2 mg/L,检测限为0.05 mg/L。     

核酸适体生物传感器快速检测牛奶中抗生素

为了现场快速检测牛奶溶液中四环素类抗生素的含量,对电极进行阳极氧化、氨基化、活化等一系列处理,将四环素核酸适体作为识别分子,共价固定于玻碳电极表面,构建出新型核酸适体生物传感器。传感器可以特异性结合标本中四环素类抗生素,并产生电化学信号,强度与抗生素浓度相关。其最低检测量是1μg/L,检测时间为5 m in。     

免标记光学纳米生物传感器的研究

利用发光稳定的多孔硅,当蛋白质溶液固定到多孔硅表面时,其发光强度和反射光谱的峰位会随着被测物质量浓度的变化而发生变化的特性。制备免标记光学纳米生物传感器来检测牛血清白蛋白（BSA）,把不同质量浓度的BSA溶液固定到多孔硅上测量反射光谱的峰位和荧光强度的变化。实验结果表明：反射光谱的峰位与牛血清白蛋白的浓度呈线性关系：y=1．4189x＋366．31,R^2=0．9898;荧光强度的降低量与BSA溶液的浓度呈线性关系,检测限为8×10^-9 mol／L。     

纳米多孔硅阻抗生物传感器的研究

构建了1种基于多孔硅材料,无需标记的纳米生物传感器,用于对牛血清白蛋白分子进行检测。通过对多孔硅进行表面处理,形成氧化膜,将抗体固定到多孔硅氧化层表面。在磷酸盐缓冲液中,通过电化学检测系统检测加入抗原后,传感器的阻抗值的变化。磷酸缓冲液（PBS）/抗体-氧化层/硅,构成电解液/绝缘层/半导体（electro-lyte-insulator-semiconductor,EIS）结构。传感器的线性检测范围为0.01～0.27mg/mL,检测限为0.01mg/mL。     

核酸适配体生物传感器对心肌肌钙蛋白Ⅰ的检测研究

核酸适配体生物传感器是利用固定在电极表面的适配子与被测溶液中心肌肌钙蛋白Ⅰ(cTnⅠ)发生特异性结合,从而达到检测的目的.我们对玻碳电极进行阳极氧化、氨基化修饰,通过碳二亚胺盐酸盐(carbodiimide hydrochloride,EDC)、N-羟基琥珀酰亚胺(N-hydroxysuccinimide,NHS)活化作用将适配子结合在电极表面.cTnⅠ最佳检测范围是0.05~5 nmol/L,最低检测限为0.05 nmol/L,检测时间为5 min.     

抗体-适配子混合夹心法检测炭疽芽孢杆菌芽孢的研究

通过SELEX(SystemEvolutionofLigandsbyEXponentialenrichment)体外筛选技术寻获的能与炭疽芽孢杆菌芽孢特异结合的DNA适配子(aptamer)考察其作为检测分子的能力。利用抗体-适配子混合夹心法,根据显色反应强度,评价适配子检测炭疽芽孢杆菌芽孢的能力。结果表明,适配子可以作为检测分子检测靶物质的存在。适配子检测范围在2-16μg,芽孢的检测范围在4×104-4×107,当适配子16μg,芽孢量为4×107显色的指示强度最强。说明适配子作为检测用分子,具有潜在的实用价值。     

适配子生物传感器对青霉素的快速检测

将玻碳电极进行阳极氧化和氨基化修饰,通过碳二亚胺盐酸盐(EDC)、N-羟基丁二酰亚胺(NHS)活化作用将青霉素适配子结合在电极表面。该适配子电化学生物传感器分子识别能力强、无放射性标记、检测速率快,青霉素类的最佳检测范围是2.81~281 nmol/L,最低检测限为2.81 nmol/L,检测时间为5 m in。     

Electrochemical aptasensor for tetracycline detection

An electrochemical aptasensor was developed for the detection of tetracycline using ssDNA aptamer that selectively binds to tetracycline as recognition element. The aptamer was highly selective for tetracycline which distinguishes minor structural changes on other tetracycline derivatives. The biotinylated ssDNA aptamer was immobilized on a streptavidin-modified screen-printed gold electrode, and the binding of tetracycline to aptamer was analyzed by cyclic voltammetry and square wave voltammetry. Our results showed that the minimum detection limit of this sensor was 10 nM to micromolar range. The aptasensor showed high selectivity for tetracycline over the other structurally related tetracycline derivatives (oxytetracycline and doxycycline) in a mixture. The aptasensor developed in this study can potentially be used for detection of tetracycline in pharmaceutical preparations, contaminated food products, and drinking water.     

Aptamer-based PDMS-gold nanoparticle composite as a platform for visual detection of biomolecules with silver enhancement

A sensitive colorimetric detection for biomolecules based on aptamer was described. Poly(dimethylsiloxane) (PDMS)-gold nanoparticles (AuNPs) composite film was used as a platform for immobilizing anti-target aptamer. PDMS-AuNPs composite film only covered with aptamer showed high inhibiting ability towards silver reduction, after target molecules were conjugated on the modified surface, the catalytic efficiency of AuNPs for silver reduction was increased. In this system, the darkness density of silver enhancement was applied for target quantitative measurement. Lysozyme and adenosine 5'-triphosphate (ATP) were tested as the models, quantitative measurements with imaging software or semiquantitative measurements with naked eyes were carried out in the range of 1×10(-2)-1 μg/mL and 1×10(-4)-1×10(3) μg/mL, the volume of reagent using in each assay is 15 μL or less. We speculated that aptamer-target conjugates' inhibition ability for AuNPs' catalytic efficiency toward silver reduction might come from charge and spatial effects. This study can offer a completely novel and relatively general approach for colorimetrical aptamer sensors with good analytical properties and potential applications. The sensor could be coupled with digital transmission of images for remote monitoring system in diagnosis, food control, and environmental analysis.     

P450-based porous silicon biosensor for arachidonic acid detection

A porous silicon biosensor based on P450 enzyme for arachidonic acid detection was developed. A new transduction method is presented with a simultaneous measurement of refractive index and fluorescence intensity changes when the analyte is binding to an enzyme on the porous silicon surface. A fluorophore bound to a cysteine residue in an allosteric position of the haem domain (BMP) of cytochrome P450 BM3 enhances its fluorescence intensity upon interaction with its substrate arachidonic acid, involved in diseases such as Alzheimer's, liver cancer and cellular inflammation processes. BMP has been anchored on porous silicon surface and the new transduction method has been successfully exploited to develop a biosensor for arachidonic acid, reaching a detection limit of 10 μM arachidonic acid in a dynamic range of 10-200 μM. Moreover, the change of the refractive index has been also monitored at the same time, displaying a higher detection limit of 30 μM. Preliminary test were also conducted in plasma proving the high specificity and selectivity of the sensor even in presence of interferents in the range of 50-100 μM. Here we suggest these two detection systems could be used simultaneously to increase the accuracy and the dynamic range of the sensor avoiding a false positive response.     

A simple assay to amplify the electrochemical signal by the aptamer based biosensor modified with CdS hollow nanospheres

A simple method to amplify the electrochemical signal by an aptamer with 22 bases modified with CdS hollow nanospheres (CdSHNs) was described. Using the thrombin as a model, the interaction between the aptamer and CdSHNs was characterized by cyclic voltammetry, electrochemical impedance spectroscopy and circular dichroism spectroscopy. CdSHNs promoted the electron transfer between the gold electrode and K(3)[Fe(CN)(6)] and facilitated the conformation conversion of the aptamer from hairpin to G-quadruplex after the aptamer interacted with thrombin. Under optimal conditions, the modified electrode could be used for the determination of thrombin from 0 to 33 μg mL(-1) and the sensitivity was 1.34 μA mL μg(-1)cm(-2), while the linear range of the modified electrode without the immobilization of CdSHNs was from 2.75 to 27.5 μg mL(-1) and the sensitivity was 0.062 μA mL μg(-1)cm(-2). This constructed biosensor also had a good stability, specificity, reproducibility and accuracy which could provide a promising platform for fabrication of aptamer based biosensors.     

Determination of glucose and uric acid with bienzyme colorimetry on microfluidic paper-based analysis devices

An aptamer is an artificial functional oligonucleic acid, which can interact with its target molecule with high affinity and specificity. Enzyme linked aptamer assay (ELAA) is developed to detect cocaine using aptamer fragment/cocaine configuration based on the affinity interaction between aptamer fragments with cocaine. The aptasensor was constructed by cleaving anticocaine aptamer into two fragments: one was assembled on a gold electrode surface, while the other was modified with biotin at 3'-end, which could be further labelled with streptavidin-horseradish peroxidase (SA-HRP). Upon binding with cocaine, the HRP-labelled aptamer fragment/cocaine complex formed on the electrode would increase the reduction current of hydroquinone (HQ) in the presence of H(2)O(2). The sensitivity and the specificity of the proposed electrochemical aptasensor were investigated by differential pulse voltammetry (DPV). The results indicated that the DPV signal change could be used to sensitively detect cocaine with the dynamic range from 0.1 μM to 50 μM and the detection limit down to 20 nM (S/N=3). The proposed aptasensor has the advantages of high sensitivity and low background current. Furthermore, a new configuration for ELAA requiring only a single aptamer sequence is constructed, which can be generalized for detecting different kinds of targets by cleaving the aptamers into two suitable segments.