Review: Recent Developments in Enzyme-Based Biosensors for Biomedical Analysis

The need for simple, rapid, cost-effective, and portable screening methods has boosted the development of practical biosensors with applications in clinical monitoring, and diagnosis of disease. Compared with traditional analytical methods, enzyme-based bioanalytical devices have several distinct advantages such as high sensitivity and specificity, portability, cost-effectiveness, and the possibilities for miniaturization and mass production. Additionally, they can be developed for point-of-care diagnostic testing. This paper reviews recent advances in the development of enzyme biosensors, design characteristics, performances, and applications with a focus on electrochemical and optical sensors. Recent emerging technologies and innovative biosensing designs, such as nanosensors, paper based-sensors, lab-on-a-chip, biochips, and microfluidic devices are discussed. Specific applications in bioanalysis, clinical diagnosis, and pharmacology are discussed.     

酶放大DNA电化学传感器的研究进展

DNA电化学传感器灵敏度高、选择性好、分析时间短和检测成本低,极大地推动了生物传感器的发展. 结合蛋白质酶、功能核酸酶的催化效率高与特异性好,可提高检测灵敏度和选择性. 本文评述了酶放大DNA电化学传感器的研究进展,并分析现存问题,展望发展趋势.     

Conducting Redoxpolymer-Based Reagentless Biosensors Using Modified PQQ-Dependent Glucose Dehydrogenase

Reagentless, oxygen-independent glucose biosensors based on an Os-complex-modified polypyrrole matrix and on soluble PQQ-dependent glucose dehydrogenase from Acinetobacter calcoaceticus are described.As the soluble form of glucose dehydrogenase from Acinetobacter calcoaceticus is a hydrophilic enzyme with a positive net charge, its entrapment into the positively charged hydrophobic polypyrrole film is much more complicated than that of the corresponding membrane enzyme or the negatively charged and very stable glucose oxidase. Possible ways for using soluble PQQ-dependent glucose dehydrogenase in combination with conducting polymer films are seen in the modulation of the enzyme properties by covalent binding of suitable compounds to the protein shell together with the adjustment of the properties of the conducting polymer film. This can be done by neutralising the net charge of the protein and/or optimising the electron-transfer pathway between enzyme and electrode surface by covalent binding of suitable redox relays to the protein surface.In addition, methods for increasing the hydrophilicity of the polymer film, such as the co-entrapment of high-molecular weight hydrophilic additives and copolymerisation of hydrophilic pyrrole derivatives are presented. It is demonstrated that the replacement of the parent monomer pyrrole by a suitable hydrophilic pyrrole derivative facilitates the entrapment of the modified soluble PQQ-dependent glucose dehydrogenase into the Os-complex-modified polymer and hence allows for the development of reagentless biosensors.     

Recent Progress on Graphene‐based Electrochemical Biosensors

The detailed records and conclusions on the important advancements in graphene‐based electrochemical biosensors have been reviewed. Due to their outstanding properties, graphene‐based materials have been widely studied for the accurate electrochemical detection of many biomolecules, which is extremely vital to the development of biomedical instruments, clinical diagnosis, and disease treatment. This review discusses the graphene research for the effective immobilization of enzymes, including glucose oxidase, horseradish peroxidase, and hemoglobin, etc., and the accurate detection of biomolecules, including glucose, hydrogen peroxide, dopamine, ascorbic acid, uric acid, nicotinamide adenine dinucleotide, DNA, RNA, and carcinoembryonic antigen, etc. In most of the cases, the graphene‐based biosensors exhibited remarkable performance with high sensitivities, wide linear detection ranges, low detection limits, and long‐term stabilities.     

Recent Advances in Electropolymerized Conducting Polymers in Amperometric Biosensors

Conducting electroactive polymers (CPs) are materials discovered just over 20 years ago which have aroused considerable interest on account of their electronic conducting properties and unique chemical and biochemical properties. Consequently, they have numerous (bio)analytical and technological applications. CPs are easily synthesized and deposited onto the conductive surface of a given substrate from monomer solutions by electrochemical polymerization with precise electrochemical control of their formation rate and thickness. Coating electrodes with CPs under mild conditions opens up enormous possibilities for the immobilization of biomolecules and bioaffinity or biorecognizing reagents, the improvement of their electrocatalytic properties, rapid electron transfer and direct communication to produce a range of analytical signals and new analytical applications. Co-immobilization of other molecules (enzymatic co-factors or charge-transfer mediators) by entrapment within electropolymerized films or by covalent binding on these films permits straightforward fabrication of reagentless biosensors. The characteristics of CPs and their uses, mainly in amperometric biosensors, are reviewed. The most recent applications and lines of research related to CP films are summarized in the different sections of the paper, and probable future trends are discussed.     

New Amperometric Cholesterol Biosensors Using Poly(ethyleneoxide) Conducting Polymers

Accumulation of cholesterol in human blood can cause several health problems such as heart disease, coronary artery disease, arteriosclerosis, hypertension, cerebral thrombosis, etc. Therefore, simple and fast cholesterol determination in blood is clinically important. In this study, two types of amperometric cholesterol biosensors were designed by physically entrapping cholesterol oxidase in conducting polymers; thiophene capped poly(ethyleneoxide)/polypyrrole (PEO-co-PPy) and 3-methylthienyl methacrylate-co-p-vinyl benzyloxy poly(ethyleneoxide)/polypyrrole (CP-co-PPy). PEO-co-PPy and CP-co-PPy were synthesized electrochemically and cholesterol oxidase was immobilized by entrapment during electropolymerization. The amperometric responses of the enzyme electrodes were measured by monitoring oxidation current of H₂O₂ at +0.7 V in the absence of a mediator. Kinetic parameters, such as Km and Imax, operational and storage stabilities, effects of pH and temperature were determined for both entrapment supports. Km values were found as 1.47 and 5.16 mM for PEO-co-PPy and CP-co-PPy enzyme electrodes, respectively. By using these Km values, it can be observed that ChOx immobilized in PEO-co-PPy shows higher affinity towards the substrate.     

生物传感器研究进展

本文详细地综述了生物传感器的类型、基本原理、目前的研究现状及应用状况,并预测了其今后的发展趋势。参考文献110篇。     

Urea Biosensors Based on PVC Membrane Ion-Selective Electrodes

Abstract

This paper presents a general method of enzyme immobilization at the surface of ion selective membranes. Covalent binding of enzymes directly on the electrode surface is a very effective method that results in stable enzymatic membranes. As an example the construction of enzymatic sensors for urea determination based on ammonium and hydrogen carbonate ion selective electrodes is presented. The optimum working conditions for these biosensors were found. Bioelectrodes based on an ammonium sensor show very good analytical parameters: dynamic stability - over 2 months without decrease of sensitivity, response time - shorter then 20 s. high sensitivity, determination range from 0.3 to 70 mM. In the contrast to the ammonium ion based biosensors, those constructed on the basis of anion selective electrodes have worse analytical parameters. It is mainly due to poor selectivity and instability of an applied ion selective electrode. In spite of this, both types of urea biosensors were used for measurements in the differential potentiometry mode. The application of such system increased the sensitivity of urea determination.     

基于局域表面等离子体共振效应的光学生物传感器*

贵金属纳米粒子表现出许多常规块体材料所不具备的优异性能,其中局域表面等离子体共振 (LSPR) 特性是研究热点之一。LSPR 的形状和位置与纳米粒子的组成、大小、形状、介电性质以及局域介质环境密切相关。基于这一特性,贵金属纳米粒子已广泛应用于光学生物传感器、光过滤器和表面增强光谱等领域。本文对各种结构的贵金属纳米粒子的制备方法及其在光学生物传感器中的应用进行了综述,并对 LSPR 纳米传感器的未来发展前景做了展望。     

Electrochemical Biosensors Based on Enzyme Inhibition Effect

Several electrochemical biosensors based on various enzyme inhibition effects have been designed; their laboratory prototypes have been manufactured and thoroughly investigated. It should be noted that such biosensors are adapted to large-scale production technologies. A number of advantages and disadvantages of developed biosensors based on enzyme inhibition has been discussed. It is important that all developed biosensors are not opposite to traditional analytical methods, but complement them. This is an additional system of quick and early warning about the presence of toxic substances in the environment. Such systems can save time and money in emergencies due to the possibility of quick decision-making on local environmental problems. If necessary, more accurate, but time-consuming and expensive traditional methods could be used for further validation and additional research of samples previously tested by biosensors.     

Capacitive Sensing of Amino Acids Using Caliraxene‐Coated Silicon Transducers

The possibility of using capacitance and flat band voltage as measurable quantities for determining amino acids that are neither electroactive nor with strong UV‐vis absorption has been explored. The sensors were fabricated by immobilizing calixarene derivatives on Si/SiO₂/Si₃N₄ transducers. The measurements were made in sulfuric acid media of ca. pH 1 and in physiological buffer of pH 7.4. The different calixarene derivatives showed varying sensitivities to the amino acids ranging from 8 to 137 mV/decade.     

生物传感器在POCT中的应用研究

即时检测（POCT）是在病人旁边或现场进行的检测,因其简单、快速、便携且不受场所限制已成为目前体外分子诊断技术发展的一支风向标。而生物传感器以其快速、灵敏、高效、便携及易于自动化、微型化等优点在发展现场即时检测技术中具有非常大的潜力。近年来,随着生物传感技术、互联网技术的发展及各种新技术、新方法的兴起和融合,POCT技术和方法得到了实质性发展。本文简要介绍了生物传感器的分类和生物传感器在POCT中的应用现状,综述了近年来各类生物传感器在面向POCT检测应用的研究进展。生物传感器根据类型主要分为基于微流控芯片的生物传感器、基于纸的生物传感器、基于纳米材料的生物传感器、基于手机检测平台的生物传感器及集成的生物传感器等,并对这些传感器平台在POCT检测方面的应用做了阐述,最后对生物传感器在POCT应用中存在的问题进行了讨论,并对其发展趋势及前景做了展望。     

有机磷农药酶生物传感器研究进展

酶生物传感器（EBS）以简单、廉价、易于微型化等优势成了有机磷农药（OPs）传统分析方法的最佳替代品。本文从识别OPs的酶及识别机理、电化学EBS、酶的固定化技术、高分子材料的酶固定载体不同角度综述了有机磷农药酶生物传感器研究近况,并重点介绍了一次性丝网印刷酶电极。     

Reversible Optical Sensor Membrane for Hydrogen Peroxide Using an Immobilized Fluorescent Probe,and its Application to a Glucose Biosensor

An optical sensor for hydrogen peroxide (HP) based on the probe europium tetracycline (EuTc) incorporated into a polyacrylonitrile-co-polyacrylamide polymer matrix is described. Upon optical excitation with 400-nm light, the EuTc in the membrane displays fairly strong fluorescence peaking at 616nm. The fluorescence intensity increases up to 3-fold if the sensor is exposed to solutions containing HP. The effect is reversible and can thus be used for continuous sensing. The largest signal changes with HP are observed at pH levels between 6.5 and 7.5, and the range of the response is between 10 and 300ppm (w/w) of HP, equal to 0.3 to 10mmolL^–1). At concentrations above 0.3% of HP, decomposition of the EuTc in the membrane is observed. The limit of detection is 15ppm (0.45mmolL^–1). The response is fully reversible and rather slow (10min) in both directions, but the reverse response may be accelerated by addition of a reducing agent such as thiosulfate. Alkali ions and most anions remain inert, but phosphate and citrate interfere, as do Cu²⁺ ions, which quench fluorescence. In order to image the spatial distribution of HP concentrations, sensor membranes were placed on the bottom of the wells of a microtiter plate, and their fluorescence was imaged using an LED-based device based on the measurement of the luminescence decay time of EuTc. If glucose oxidase is immobilized on the sensor layer, a glucose sensor is obtained in which the HP sensor acts as the transducer and which can quantify glucose in concentrations between 0.1 and 5mmolL^–1. The limit of detection for glucose is 0.2mmolL^–1.     

纳米粒子在电化学DNA生物传感器研究中的应用

简要介绍了电化学DNA生物传感器的原理和分类,对纳米粒子在电化学DNA生物传感器研究中的应用进行了详细评述.     

Lead Sensor Using Gold Nanostructured Screen‐Printed Carbon Electrodes as Transducers

Gold nanostructured screen‐printed carbon electrodes are demonstrated to be suitable transducers for the determination of lead using square‐wave voltammetry. Reproducible gold nanostructures have been obtained by direct electrochemical deposition. A calibration plot from 2.5 to 250 μg/L was obtained in acidic solutions of Pb(II) with a reproducibility of 4% (n=10). The detection limit was 0.09 μg/L of lead. The method is then applied to perform a blood lead analysis by adjusting square‐wave parameters in capillary or venous blood with a minimum sample pretreatment and excellent accuracy and reproducibility.     

石墨烯纳米复合材料在电化学生物传感器中的应用

将石墨烯与其他纳米材料复合,是一种拓展或增强其应用的有效方法。借助不同组分间的协同作用,可以改善石墨烯的电学、化学和电化学性质,拓展和增强石墨烯的电化学效应,为固定氧化还原酶,实现直接电化学提供新型、高效的平台,应用于第三代电化学生物传感器的设计和制备,对葡萄糖、胆固醇、血红蛋白、DNA、H₂O₂、O₂、小生物分子等的检测显示出了优异的灵敏度和选择性。本文综述了基于石墨烯构筑的纳米复合材料在电化学生物传感器中的应用研究,包括石墨烯与贵金属、金属氧化物/半导体纳米粒子、高分子、染料分子、离子液体、生物分子等的纳米复合材料,并对石墨烯材料在电化学领域的发展方向和应用前景进行了展望。     

偶氮液晶在光学材料中的应用进展

偶氮液晶是一类分子中含有偶氮苯结构的液晶,其偶氮基团在光照射下,能发生可逆的顺反异构反应。由于偶氮苯的顺式结构不能形成稳定的液晶相,因此,偶氮液晶在具有向列液晶温敏性质的同时,也具有特有的光敏性质,其相变与分子取向能够通过光照进行控制。这种特性使得偶氮液晶在光学功能材料方面具有广泛的应用。本文介绍了偶氮液晶的基本特性,结合作者自身的研究工作阐述了其在光学材料应用方面的研究进展。     

基于散射信号的超分辨光学相减显微镜

现有的光学超分辨显微成像技术主要依赖于特殊的荧光标记物,其对于大多数非荧光样品的超分辨成像就变得无能为力。因此我们提出将光学相减显微技术应用到非荧光样品的成像当中,利用普通共聚焦光斑和面包圈型光斑分别激发样品的散射光成像,从而得到样品同一区域的两幅图像,再通过图像相减的方法提高了图像空间分辨率。不同于一般的超分辨成像方法,这种光学相减显微镜不需要特殊的样品预处理过程,同时两次成像的激发光强度可以保持在一个较低水平,避免了样品损伤的影响。随后金纳米小球和有机聚合物微丝的散射成像实验证明了光学相减显微镜可以将空间分辨率提高到215 nm (0.33λ, 1λ = 650 nm),并且通过探测散射信号得到更多的样品细节信息。     

葡萄糖生物传感器检测方法的研究进展

葡萄糖生物传感器以其灵敏度高、选择性好、反应速度快以及稳定性好等优点吸引了许多研究者的关注。 本文将已发表的一些葡萄糖检测方法分为两类：葡萄糖酶生物传感器检测方法与无酶葡萄糖生物传感器检测方法,简要介绍了这2种检测方法的一些研究进展,并对葡萄糖检测方法的发展前景进行了展望。