碲化钴纳米管/纳米金/壳聚糖复合膜DNA电化学传感器的构建

通过水热法制备了碲化钴纳米管,以制备的碲化钴纳米管、纳米金和壳聚糖组成的复合膜修饰裸铂电极,构建了一新型DNA电化学生物传感器.利用该生物传感器及邻菲罗啉钴电化学杂交指示剂对禽病毒基因进行了检测研究.实验表明,该法测定DNA的浓度线性范围为2.0×10-10～2.0×10-6 mol/L,最低检测限为7.94× 10-11 mol/L.     

基于纳米金增强吸附伏安分析的电化学免疫传感技术用于蛋白分子检测

基于纳米金标记和金增强表面吸附伏安分析的新型电化学免疫传感器.以人免疫球蛋白G(h IgG)为模型分析物,使固定化抗体与分析目标物hIgG及纳米金标记的hIgG抗体结合于电极表面,通过金沉积使电极表面的纳米金颗粒直径增大.由于金增强时间在几分钟内就可完成,和传统的银增强相比,缩短了增强时间,加快了分析速度.研究结果表明,该法灵敏度高,选择性好,重现性好,操作简便,可望成为一种有潜力的新型高灵敏度免疫传感技术.     

结合PAH聚电解质和纳米金界面的压电免疫传感器研究

提出了一种聚丙烯氯化铵(PAH)-纳米金固定抗体的压电免疫传感器界面的构建方法.先在压电石英晶振的金电极表面自组装一层半胱氨酸单层膜,通过戊二醛交联带大量NH2基的聚电解质PAH,随后在PAH膜表面自组装一层纳米金粒子,以静电吸附作用固定IgG抗体,研制成一种新的压电免疫传感器的界面,用于对相应抗原的检测.研究了PAH浓度及抗体固定化等实验条件的影响,探讨了传感器的主要响应特性与再生性能,并与戊二醛直接固定的传感器的性能进行了比较.结果表明,前者固定的抗体的活性较高,响应频率较大,检测的线性范围较宽,非特异性吸附小,能有效地改善传感器的灵敏度和检测限,而且容易进行传感器的再生.     

基于纳米金颗粒放大的电化学传感器用于三聚氰胺的检测

三聚氰胺与胸腺嘧啶（T）之间能够通过三个氢键结合,以富T的DNA探针为识别元件,结合DNA修饰的纳米金颗粒放大技术,以电活性物质钌胺作为信号分子,发展了一种高灵敏检测三聚氰胺的电化学传感器,该传感器具有良好的特异性和灵敏度,检测下限低至0．5nmol／L。     

电化学/生物传感器快速检测大肠杆菌的研究进展

大肠杆菌广泛分布于自然界中,通常被用来作为水体系统排泄物污染情况的指示菌.它是大面积食物中毒的主要原因之一,严重感染者会引发败血症、肾功能衰竭等危及生命的并发症.电化学/生物传感器具有独特的优势,如能在浑浊溶液中操作、选择性好、灵敏度高、检测速度快等,因此在临床检测、环境保护和食品安全等领域得到了广泛应用.该文主要对电化学,生物传感器快速检测大肠杆菌的研究进展进行了简要的综述.     

纳米金结合银增强用于质量放大压电免疫传感器的设计

提出了一种基于纳米金结合银增强的质量放大方法以提高压电免疫传感器的灵敏度.通过夹心法将纳米金引入传感器表面后滴加银增强试剂,在纳米金的催化下可使银沉积在传感器上,从而对免疫反应引起的质量变化进行放大.以检测正常人IgG为例,对该方法的可行性作了研究,设计的传感器可在0.3～5μg/mL范围内实现对正常人IgG的定量检测.     

基于纳米金/硫堇修饰金电极的脱落酸安培免疫传感器的研制

提出了一种基于纳米金/硫堇修饰金电极的ABA安培免疫传感器。该传感器基于H2O2-HRP-硫堇催化波体系构建,其中硫堇为传感介质。当HRP存在时,通过加入H2O2,硫堇的还原电流大幅增加,并且电流的增加依赖于HRP活性。HRP活性又由ABA与HRP酶标抗体结合物调控,产生一个减小的催化波。用BSA封闭硫堇单分子层修饰后可能存在的活性位点以避免非特异性吸附。优化了测定条件,包括酶标抗体和硫堇的最佳比例、培育时间、缓冲液的pH值和H2O2浓度。此传感器的还原电流在ABA浓度0.5~1000ng/mL范围内呈线性下降,回归方程为y=0.0209x 17.071,相关系数为0.9922,检测限为0.2ng/mL。     

快速检测大肠杆菌O157:H7的电化学阻抗免疫生物传感器

建立了一种采用电化学阻抗谱技术快速检测大肠杆菌O157∶H7的生物传感器,它是通过石英晶体金电极表面附着一层蛋白A膜来固定抗体的。该生物传感器采用了三电极系统-工作电极石英晶体金电极、Ag/AgCl/Cl-SAT参考电极和铂对电极。和以往的普通金电极不同,第一次采用了石英晶体金电极。试验结果说明抗体的固定以及大肠杆菌O157∶H7与抗体的结合都增加了石英晶体金电极表面的电子传递阻抗,在[Fe(CN)6]3-/4-氧化还原对存在的情况下,用电化学阻抗谱测量该阻抗。该免疫生物传感器的检测限是103cfu/mL,石英晶体金电极的电子传递阻抗变化值和大肠杆菌O157∶H7的浓度在一定范围内呈线性关系,检测时间少于10 min。     

基于电聚合硫堇膜与纳米复合材料界面的直接电化学免疫传感器的构建

本工作采用电聚合手段在玻碳电极表面制备聚硫堇基质膜,借硫堇膜上富含的氨基基团实现对碳纳米管与纳米金颗粒二元纳米复合材料的化学组装,制备稳定的碳纳米管/纳米金/聚硫堇传感界面, 以此传感界面固定免疫活性物质,发展了一种无电子媒介的酶免疫传感器.结果表明,电极采用碳纳米管/纳米金颗粒界面固定免疫活性物质,其传感响应性能明显优于单独采用碳纳米管的情况.以人IgG免疫体系为例,电极以此界面固定人IgG抗体,并通过夹心方式直接响应酶催化H2O2的还原电流信号,实现了对人IgG浓度的灵敏测定.此外,采用甘氨酸-HCl缓冲液洗涤使用后的传感器,可有效洗脱其表面的免疫复合物,实现传感器的重复使用.     

检测大肠杆菌O157:H7的电化学阻抗谱生物传感器的研究

我们提出了用掺锡的三氧化二铟(ITO)作为工作电极,通过硅烷化固定化技术,将抗大肠杆菌O157:H7单克隆抗体固定在ITO电极表面,利用电化学阻抗谱技术来构建一种新型的免疫传感器.该新型的免疫传感器的检测限为 4×103CFU/mL,检测线性范围为4×103-4×106CFU/mL.实验研究表明,该传感器具有灵敏度较高,检测时间短,操作简单等优点,在临床医学和环境监测中具有应用价值.     

基于Nafion-石墨烯复合物固定甲胎蛋白电化学免疫传感器的研究

该文利用Nation-石墨烯复合物和纳米金固定甲胎蛋白抗体(anti-AFP),构建了高灵敏的电化学免疫传感器.首先将石墨烯分散在Nation溶液中制得Nation-石墨烯的复合膜,并将其固定在玻碳电极(GCE)表面,通过静电吸附和共价键合作用将硫堇(Thi)和纳米金颗粒(nano-Au)依次固定到Nation-石墨烯复合膜修饰的玻碳电极表面.再通过纳米金单层吸附anti-AFP,最后用牛血清蛋白(BSA)封闭电极上的非特异性吸附位点,从而制得了甲胎蛋白免疫传感器.实验结果表明,该修饰电极对不同浓度的甲胎蛋白(AFP)有很好的响应,其线性范围为0.8～100 ng/mL,检出限为0.36 ng/mL.     

Development of an immunosensor based on the measurement of fluorescence

Consumers concern about the presence of pesticides in food is increasing. However current techniques for detecting pesticide residues in olive oil are very expensive and slow, which make them difficult to apply in commercial conditions. For this reason, the Instituto Valenciano de Investigaciones Agrarias (IVIA) and Universidad Politécnica de Valencia (UPV) have decided to develop an immunosensor as an alternative, fast and cheap method to measure the concentration of some pesticides in olive oil. This work shows the developed transducer by IVIA based on a photovoltaic sensor that transforms the fluorescent radiation to an electrical signal. The immunosensor is based on an immunologic reaction which causes fluorescence on the sample. The intensity of the fluorescence is related with the concentration of marked antibodies. The performed tests with two types of immunoassays and fluorescent markers (FITC and ELF97) have demonstrated the robustness and reliability of the developed transducer. The immunosensor is able to discriminate different concentrations of marked antibodies. This study has demonstrated the possibility of applying this immunosensor to detect chemical residues from pesticide treatments in olive oil samples. These results validate the sensor for future commercial applications.     

Label-free electrochemical immunosensor for sensitive detection of kanamycin

A novel label-free electrochemical immunosensor for sensitive detection of kanamycin based on water-soluble graphene sheet (WGS)/prussian blue-chitosan (PB-CTS)/nanoporous gold (NPG) composited film has been reported. PB was selected as an electron transfer mediator, and was modified onto the electrode together with WGS through electrostatic adsorption. Then NPG was immobilized onto the as-prepared film for biomolecules anchoring. The electroactivity of PB was greatly enhanced in the presence of WGS and NPG. It could mainly be ascribed to the fact that the good conductivity of WGS and NPG promoted electron transfer and enhanced the sensitivity. kanamycin antibody, as a model, was immobilized onto the composite film for the detection of kanamycin. Under optimum conditions, the amperometric signal of PB decreased linearly with kanamycin concentration (0.02-14 ng mL⁻¹), a linear calibration plot (y = 1.3817 + 4.7544x, r = 0.9993), resulting in a low limit of detection (6.31 pg mL⁻¹). The novel immunosensor for the detection of kanamycin in real sample with satisfactory results has been proved. In addition, this method would be easily adapted for the detection of other residual antibiotics in animal derived foods.     

Sensitive electrochemical immunosensor based on enlarged and surface charged gold nanoparticles mediated electron transfer

A simple and sensitive electrochemical immunosensor for the detection of protein biomarker interleukin-6 (IL-6) based on gold nanoparticles (AuNPs) as label was reported. The signal of the immunosensor was originated from enlarged and positively charged AuNPs mediated electron transfer between insulating self-assemble monolayer (SAM) modified electrode and K₃Fe(CN)₆ solution. The gold electrode was first modified with SAM to block the electron transfer between the electrode and K₃Fe(CN)₆ solution. After the preparation of the immunosensor, the AuNPs attached onto electrode was enlarged and positively charged when treated in solution containing HAuCl₄, ascorbic acid and cetyltrimethylammonium bromide (CTAB). The enlarged and positively charged AuNPs then mediated electron transfer between the electrode and K₃Fe(CN)₆ solution, creating redox current that is proportional to the concentration of IL-6 detected. The reported immunosensor has high sensitivity and wide linear range. This novel signal amplification strategy can be applied to the detection of different kinds of protein biomarkers.     

基于核酸识体的电化学免疫传感器用于血小板源性生长因子的检测

该文提出了一种新颖的、高灵敏度的基于核酸识体和银沉积的电化学免疫传感器用于人体血小板源生长因子BB(PDGF-BB)的检测.首先在电极的表面固定抗体,然后与抗原、生物素标记的核酸识体形成免疫夹心复合物,再利用生物素和亲合素的特异性反应把亲合素标记的碱性磷酸酶固定到电极上,然后把得到的产物在底物溶液中进行银沉积,最后利用电化学方法对沉积到电极表面的银进行检测以达到检测PDGF-BB的目的.实验对固定抗体浓度和生物素标记核酸识体的浓度两个条件进行了优化.实验结果表明PDGF-BB的浓度在1～1000 ng/mL的范围具有良好的线性关系,最低检测下限可达0.8 ng/mL.该方法具有高灵敏度和稳定性,在临床上具有广阔的应用前景.     

基于纳米材料的电化学生物传感器研究进展

该文主要从纳米尺寸材料电极的构建以及纳米材料作为生物分子指示剂两部分展开讨论,描述了依赖于阵列纳米管排列的生物分子与电极间的直接电子传递,纳米管、纳米颗粒为基质的纳米电极的构建,以及以金纳米颗粒、DNA量子点和蛋白为基础的多路分析技术与负载于CNT的新的纳米生物标记,特别讨论了纳米电化学方法在检测DNA和免疫传感器领域取得的研究进展.     

Scores of generalized base properties for quantitative sequence-activity modelings for E. coli promoters based on support vector machine

A novel base sequence representation technique, namely SGBP (scores of generalized base properties), was derived from principal component analysis of a matrix of 1209 property parameters including 0D, 1D, 2D and 3D information for five bases such as A, C, G, T and U. It was then employed to represent sequence structures of E. coli promoters. Variables which were used as inputs of partial least square (PLS) and support vector machine (SVM) were selected by genetic arithmetic-partial least square. All samples were divided into train set which was applied to develop quantitative sequence-activity modelings (QSAMs) and test set which was used to validate the predictive power of the resulting models according to D-optimal design. Investigation on QSAM by PLS showed properties of base of position -42, -34, -31, -33, -41, -46 and -29 may yield more influence on strengths, which has thus pointed us further into the direction of strong promoters. Parameters of SVM were determined by response surface methodology. Satisfactory results indicated that the simulative and the predictive abilities for the internal and external samples of QSAM by SVM were better than those of PLS. Those results showed that SGBP is a useful structural representation methodology in QSAMs due to its many advantages including plentiful structural information, easy manipulation, and high characterization competence. Moreover, SGBP-GA-SVM route for sequences design and activities prediction of DNA or RNA can further be applied.     

一种基于纳米金／石墨烯／普鲁士蓝（PB）修饰玻碳电极非标记免疫传感器

采用电聚合的方法将普鲁士蓝（PBl聚合在玻碳电极表面,再将石墨烯修饰在PB上面,然后再采用电沉积的方法将HAuCL直接还原成纳米金粒子,沉积在石墨烯表面,最后将羊抗人IgG抗体直接固定于该修饰的玻碳电极表面,制备了用于人IgG抗原检测的非标记电化学免疫传感器。利用循环伏安法和交流阻抗研究了修饰电极表面的电化学特性,用差分脉冲伏安法对人IgG抗原进行了测定。实验表明,此免疫传感器在含不同浓度人IgG的PBS溶液（pH6．98）中测定,响应电流与人IgG浓度在5．55～455．5ng／mL范围内有良好的线性关系,其相关系数r=0．9926,检测限为0．015ng／mL（S／N=3）。该免疫传感器具有制备简单、响应时间快（5min）、稳定性好等特点。     

Development of sulfhydrylated antibody functionalized microcantilever immunosensor for taxol

An immobilization method using a sulfhydrylated monoclonal antibody (mAb) on the gold surface of a microcantilever immunosensor was developed for label-free detection of taxol. The sulfhydrylated anti-taxol mAb was synthesized. Then it was immobilized on the gold surface of the microcantilever. The deflection of the microcantilever corresponding to different taxol concentrations was real time monitored by optical lever technique. The activity of the anti-taxol mAb before and after sulfhydrylation, and the binding of the sulfhydrylated anti-taxol mAb on the gold surface of the microcantilever were evaluated by non-competitive enzyme-linked immunosorbent assay (ELISA). The limit of detection is 1 ng/mL and the method was used for detection of taxol in human plasma sample.