电致化学发光生物检测技术的新进展

电致化学发光作为一种分析技术,不仅可用于化学分析,而且正在被越来越多地用于生物检测和传感技术中。随着该分析技术与免疫检测技术生化固定化技术和微细加工技术等的相互融合,电致化学发光生物检测技术具有了更高的精度、分辨率和更广的应用范围。     

电致化学发光研究的新进展

综述了近１０年来国内外电致化学发光（ＥＣＬ）在分析化学上的研究及应用情况。根据反应类型对ＥＣＬ进行了分类，阐述了它们的反应机理，并展望了ＥＣＬ的发展趋势。     

鲁米诺电致化学发光机理的研究

电致化学发光（简称电致发光）是某些化学物质电解过程伴随光产生的现象·鲁米诺（Lummol，3一氨基苯一甲酸讲）在碱性水溶液是一种典型的电致发光材料·Havey，Kuwana和H。即把k。等人［’－‘］分别在二十、六十和／＼十年代对其发光进行过研究．他们都采用共存于一室的电极体系同时实现阳、阴极电解得到电致发光．HaaPakka等采用旋转环盘电极研究其发光机理．他们在盘电极上维持一定的负电位，在环电极上施加对称双阶跃电位．因此，他们认为发光过程是盘电极首先将分子氧还原成过氧化氢，通过电极的旋转，盘上产物被输送到环电极附近…     

ADPD电致化学发光法测定稀土元素

本文在自制的ECL-1型电致化学发光仪上,用电致化学发光分析法研究了稀土元素钇(Ⅲ)催化新试剂5-(对-苯胺偶氮)-2,3-二氢-1,4-酞嗪二酮(简称ADPD)的电致化学发光行为。发现Y(Ⅲ)催化ADPD电化学发光具有较好的选择性,实验了23种阳离子,只有Ag~+、Ni~(2+)、La(Ⅲ)、Cu~(2+)、Sc(Ⅲ)等干扰比较严重。本法可望发展为单项测定钇的新方法。     

电致化学发光研究的新材料和新方法

由于方法的使用范围广、光学系统简单和操作容易,电致化学发光(ECL)得到人们的广泛重视。随着对ECL研究的深入,ECL研究所涉及的领域和层面已有很大的扩展,特别是近十年来,ECL研究发展更为迅猛。除ECL理论研究外,为了适应分析检测的应用的需求,ECL在新材料、新实验技术和方法方面出现了许多的研究报道。本文综述最近几年来ECL研究在新材料应用和新实验技术的开发方面的一些进展,包括纳微米材料和量子点材料在ECL方面的研究,同时对固态ECL和基于三原色(RGB)机理的可视化ECL研究进展,进行了一些讨论。最后,综述展望纳米和量子点材料修饰电极ECL的研究和应用的前景。     

铱配合物的电致化学发光性能研究

结构相关的铱配合物[Ir(ppy)2L1](PF6)和[Ir(ppy)2L2](PF6)(ppy=2-苯基吡啶,LI=4-(2,2'-联吡啶-3-乙炔基)-N-(吡啶-2-亚甲基)-N-(噻吩-3-亚甲基)苯胺,L2=4-(2,2'-联吡-3-乙炔基)-N-二(吡啶-2-亚甲基)-苯胺)都具有优良的电致发光(ECL)性...     

碱性水溶液中ABEI的电致化学发光的研究

研究了6-[N-(4-氨基丁基-N-乙基)-N-乙基]-氨基-2,3-二氢吩噻嗪1,4-二酮(ABEI)的电致化学发光的各种条件,发现最佳的电脉冲参数为:占空比0.45,脉冲周期20ms,脉冲幅值+1.6V(相对饱和甘汞电极);最佳介质是0.12mol/L KOH-0.080mol/L H_3BO_(3-)0.080 mol/L KCI(pH12.0).在这些条件下,ABEI的发光强度与浓度在1.0×10~(-8)～9.0×10~(-5)mol/L范围内呈线性关系.研究表明,溶液中的Cl~-先被氧化为CIO~-,CIO~-再与ABEI作用并使其发光.     

电致化学发光分析法测定苯酚的研究

电致化学发光是指通过电化学方法在电极表面产生一些特殊的物质,这些物质之间或与体系中其它组分之间通过电子传递形成激发态,由激发态返回到基态产生发光现象.     

罗丹明B的电致化学发光行为研究

研究发现罗丹明B在碱性溶液中铂电极上有较强的电致化学发光行为.通过对不同NaOH浓度,以及对不同支持电解质的考察,确定最佳电致化学发光条件.在最优条件下,在1.2×10-7~1.1×10-6mol/L浓度范围内,罗丹明B的电致化学发光强度与其浓度成线性关系,最低检测限为9.0×10-8mol/L(S/N=3).将罗丹明B同一些生物活性物质相配合,然后通过罗丹明B的ECL技术对生物活性物质进行检测.     

毛细管电泳-电致化学发光测定诺氟沙星的研究

基于诺氟沙星对联吡啶钌在铂电极上的电致发光信号有增敏作用，与毛细管电泳结合，建立了一种测定诺氟沙星的电化学发光分析新方法。研究了工作电极电位、磷酸盐缓冲液浓度及其pH值、进样电压和进样时间等实验参数对诺氟沙星测定的影响。在优化的实验条件下，其浓度线性范围为0．02～10μmol／L；检出限（3σ）为0.0048μmol／L；峰面积的相对标准偏差为2．6％（1．0μmol／L，n=11）。本法可直接用于尿液中诺氟沙星（NFLX）含量的测定。回收率为92．7％～97．9％，结果满意。     

基于DNA电化学发光传感器研究金纳米颗粒对量子点的电化学发光影响

纳米金颗粒具有高的消光系数和良好的表面等离子体共振特性, 其等离子体共振特性受纳米金颗粒的尺寸和周围环境等因素的影响. 本文基于半导体纳米晶电化学发光信号对金纳米颗粒的距离依赖性制备了DNA电化学发光传感器. 首先利用循环伏安法(CV)在玻碳电极(GCE)表面原位沉积金纳米颗粒(AuNPs), 巯基丙酸包裹的CdS量子点(QDs)与氨基修饰的双链DNA (dsDNA)通过酰胺键缩合, 形成量子点修饰的双链DNA(QDs-dsDNA). 最后将QDs-dsDNA 通过dsDNA 另一端的巯基组装到纳米金表面, 得到CdS QDs-DNA/AuNPs/GCE电化学发光传感器. 在优化电极表面QDs-dsDNA密度、金纳米颗粒沉积方法等实验条件的基础上, 对不同传感器的表面性质进行了表征, 如形貌和电化学阻抗等. 进一步通过控制纳米金和CdS QDs之间的DNA研究了纳米金对CdS QDs发光信号的影响作用. 结果显示DNA链的长度和类型对发光信号有着重要的影响. 最后将此传感器用于环境污染物的DNA损伤检测, 显示出很好的灵敏响应.     

Engineering Micrometer-Sized DNA Tracks for High-Speed DNA Synthesis and Biosensing

φ29 DNA polymerase (Polφ29) is capable of synthesizing long-chain single-stranded (ss) DNA molecules by copying the sequence of a small ss circular DNA template (ssCDT) in a process known as rolling circle amplification (RCA). The use of a ssCDT in RCA, however, comes with a key drawback: the rate of DNA synthesis is significantly reduced. We hypothesize that this issue can be overcome using a very long linear ssDNA template with a repeating sequence. To test this idea, we engineered a DNA assembly, which we denote “micrometer-sized DNA track” (μDT). This μDT, with an average length of ≈13.5 μm, is made of a long chain DNA with a primer-binding domain at its 3′ end and ≈1000 repeating sequence units at its 5′ end, each carrying a DNA anchor. We find that Polφ29 copies μDT at a speed ≈5-time faster than it does a related ssCDT. We use this to design a simple all-in-one printed paper device for rapid and sensitive detection of microRNA let-7. This paper sensor is capable of detecting 1 pM let-7a in 10 minutes.     

DNA水凝胶在生物传感中的应用和发展

脱氧核糖核酸(DNA)是一种重要的生物分子,具有许多独特的性质如:信息传递、分子识别、可编辑等.DNA水凝胶同时具有DNA分子和水凝胶材料的优势,并且可以引入其他纳米材料获得多功能杂化水凝胶.相比于传统水凝胶,DNA水凝胶具有良好的特异识别能力以及可以按需设计的性质,从而被广泛应用于生物传感领域.本文围绕DNA水凝胶的...     

Investigation of DNA Pesticide Interactions by Sensitive Electrochemiluminescence Method

A solid-state [Ru(bpy)₂(dppz)]²⁺ (bpy = 2,2′-bipyridine, dppz = dipyrido[3,2-a: 2′,3′-c]phenazine) electrochemiluminescence (ECL) biosensor for studying the binding interactions between pesticides of heterocyclic polycyclic aromatic hydrocarbon (heteroPAH) and natural double-stranded DNA (ds-DNA) was constructed. Layer-by-layer films of negatively charged natural ds-DNA and polycationic poly (diallyldimethylammonium chloride) (PDDA) were assembled on the surface of a glassy carbon electrode (GCE). The complex of [Ru(bpy)₂(dppz)]²⁺ was used as a probe. Tripropylamine (TPA) was used as an electron donor to chemically amplify the ECL intensity of the probe. If the xenobiotic molecules compete with the probe for the same site on the DNA film, it would displace the probe from the DNA to decrease the ECL signal. The interactions of DNA with three pesticide molecules, quinalphos, quinclorac and carbendazim, were studied. From the displacement curve, the values of binding constant K _b of three pesticides to DNA is determined, which is in the range of 0.5 × 10⁴ to 2.3 × 10⁴ M^?1.     

硫化镉量子点-纳米金复合颗粒电化学发光传感器研究DNA的损伤

采用电化学发光方法研究了全氟辛酸(Perfluorooctanoic acid,PFOA)对DNA的损伤。结合量子点(Quantum dots,QDs)及纳米金(Nano gold,NG)颗粒的独特性能,制备了量子点-纳米金复合颗粒。将小牛胸腺DNA(ct-DNA)修饰在玻碳电极表面,然后修饰量子点-纳米金复合颗粒,构建了基于纳米金的量子点电化学发光传感器,研究了纳米金对量子点发光强度的增强作用,并利用该传感器进一步研究了PFOA对ctDNA的损伤作用。采用原子力显微镜(AFM)及X射线光电子能谱(XPS)技术对修饰电极的表面形态进行表征。实验结果表明,与单一的量子点电化学发光传感器相比,纳米金-量子点复合物电化学发光传感器的发光强度增大了近4倍。同时,ct-DNA经PFOA温浴作用后,电化学发光强度发生显著降低,表明PFOA导致了ct-DNA损伤。     

Programmable DNA Nanoflowers for Biosensing,Bioimaging, and Therapeutics

DNA nanostructures have shown excellent prospects in biomedical applications owing to their unique sequence programmability, function designability, and biocompatibility. As a type of unique DNA–inorganic hybrid nanostructures, DNA nanoflowers (DNFs) have attracted considerable attention in the past few years. Precise design of the DNA sequence enables the functions of DNFs to be customized. Specifically, DNFs exhibit high physiological stability and more diverse properties by virtue of the incorporation of inorganic materials, which in turn have been applied in an assortment of biomedical fields. In this review, the design, synthesis, and biomedical applications of programmable DNFs are discussed. First, the background of DNA-based materials and the fundamentals of DNFs are briefly introduced. In the second part, two synthetic methods of DNFs are categorized as the rolling circle amplification and salt aging method, focusing on the formation mechanism of DNFs and differences between the synthetic methods. In the third part, the biomedical applications of DNFs functional materials are summarized, including biosensing, bioimaging, and therapeutics. Finally, the challenges and future opportunities of DNFs are discussed toward more widespread applications.     

In Vitro Selection of Circular DNA Aptamers for Biosensing Applications

We report on the first effort to select DNA aptamers from a circular DNA library, which resulted in the discovery of two high‐affinity circular DNA aptamers that recognize the glutamate dehydrogenase (GDH) from Clostridium difficile, an established antigen for diagnosing Clostridium difficile infection (CDI). One aptamer binds effectively in both the circular and linear forms, the other is functional only in the circular configuration. Interestingly, these two aptamers recognize different epitopes on GDH, demonstrating the advantage of selecting aptamers from circular DNA libraries. A sensitive diagnostic test was developed to take advantage of the high stability of circular DNA aptamers in biological samples and their compatibility with rolling circle amplification. This test is capable of identifying patients with active CDI using stool samples. This work represents a significant step forward towards demonstrating the practical utility of DNA aptamers in clinical diagnosis.     

Batch Injection Analysis with Amperometric Detection for DNA Biosensing Applications

In this work, batch injection analysis with the amperometric detection (BIA‐AD), employing a detection cell designed to adapt a screen‐printed carbon electrode (SPCE) was used for the first time as a robust electroanalytical system for DNA biosensing applications. The sensitive amperometric detection was used to evaluate the structural changes in double‐stranded DNA (dsDNA) after UV‐C irradiation of its solution for a given time. Batching of DNA samples was performed by precise electronic pipette microinjection of an irradiated sample aliquot onto the unmodified activated SPCE surface incorporated in the BIA‐AD system. Using the optimized experimental conditions (40 μL of 1 mg mL^?1 dsDNA in a 0.1 M phosphate buffer of pH 7.4 sampled at the injection speed degree of 6 and detected at the potential of +1.5 V vs silver pseudo‐reference electrode), a time‐dependent response (gradual decrease of amperometric signal up to 58 % after 10 min of the irradiation) was found for the detection of damage to low molecular weight salmon sperm dsDNA. The advantages of this low‐dimensional and cost‐effective measuring system can be utilized not only for the quantification of DNA damage/degradation by UV irradiation, but they are also promising for studying other types of DNA interactions.     

Programmable Engineering of a Biosensing Interface with Tetrahedral DNA Nanostructures for Ultrasensitive DNA Detection

Self‐assembled DNA nanostructures with precise sizes allow a programmable “soft lithography” approach to engineer the interface of electrochemical DNA sensors. By using millimeter‐sized gold electrodes modified with several types of tetrahedral DNA nanostructures (TDNs) of different sizes, both the kinetics and thermodynamics of DNA hybridization were profoundly affected. Because each DNA probe is anchored on an individual TDN, its lateral spacing and interactions are finely tuned by the TDN size. By simply varying the size of the TDNs, the hybridization time was decreased and the hybridization efficiency was increased. More significantly, the detection limit for DNA detection was tuned over four orders of magnitude with differentially nanostructured electrodes, and achieved attomolar sensitivity with polymeric enzyme amplification.