固定化蛋白质在沸石和介孔分子筛上的直接电子转移及生物传感

氧化还原蛋白质和电极之间的直接电子转移在理论和实验上都具有重要的意义.通过对蛋白质和电极的修饰能够促进它们之间的直接电子转移.蛋白质与电极表面直接接触易导致蛋白质变性,在电极表面固定沸石或介孔分子筛可为氧化还原蛋白质提供一个类似于天然状态的微环境,通过分子筛的传输通道加速直接电子转移.本文主要介绍在沸石和介孔分子筛上固定化的蛋白质的电子转移和生物传感,并重点评述这些方面的研究进展、遇到的挑战及发展趋势.     

直接电化学酶传感器的研究进展

电化学生物传感器是电化学、电分析化学以及生物学等多学科交叉的产物,是一种全新的检测仪器,具有快速、灵敏和价廉等特点,在食品发酵工业、临床医学、环境监测、军事等领域有着重要的应用价值.简述了电化学生物传感器的基本原理和分类、电流型生物传感器的发展和特点以及直接电子转移的基本原理,并深入探讨了基于纳米材料等各种新型功能材料的直接电化学酶生物传感器的研究进展.     

直接电化学酶传感器的研究进展

电化学生物传感器是电化学、电分析化学以及生物学等多学科交叉的产物,是一种全新的检测仪器,具有快速、灵敏和价廉等特点,在食品发酵工业、临床医学、环境监测、军事等领域有着重要的应用价值。简述了电化学生物传感器的基本原理和分类、电流型生物传感器的发展和特点以及直接电子转移的基本原理,并深八探讨了基于纳米材料等各种新型功能材料的直接电化学酶生物传感器的研究进展。     

一步电化学还原氧化石墨烯-壳聚糖复合膜固定的辣根过氧化物酶的直接电化学及对过氧化氢的传感检测

为构筑出一种新的辣根过氧化物酶(HRP)第三代电化学生物传感器并将其用于H_2O_2的有效检测,采用循环伏安法将滴涂于玻碳电极(GCE)表面的壳聚糖(CS)-氧化石墨烯(GO)复合膜一步还原成壳聚糖(CS)-电化学还原氧化石墨烯(ErGO)复合膜,然后结合一层CS-辣根过氧化物酶(HRP)复合物,制备出CS-HRP/CS-ErGO/GCE,其中内层CS用于吸附HRP,外层CS用于阻止HRP泄漏。利用复合膜中Er GO良好的导电性和电催化性能,实现HRP与电极表面的直接电子转移。此外,CS/CS-ErGO还为HRP提供一个生物相容性微环境,使得修饰在电极上的HRP能保持其生物活性。结果表明:该修饰电极在空白磷酸盐缓冲液(PBS)溶液中出现一对氧化还原峰,式量电位为-0.11 V(vs.Ag/Ag Cl),说明包埋在CS/CS-ErGO膜中的HRP与玻碳电极之间发生了直接电化学行为。此外,该修饰电极对H_2O_2的还原具有电催化作用,能快速、灵敏地响应H_2O_2的浓度变化,其线性范围为1.0×10~(-5)~7.0×10~(-4)mol/L,检测限为3.0×10~(-6)mol/L(3S/N)。该传感器具有制备方法简单、成本低廉且稳定性良好的特点。     

血红蛋白在银微粒修饰镍铬电极上的直接电化学行为研究

以镍铬合金为基体电极,采用电沉积技术制备了银微粒电极.用扫描电镜表征了基体电极电沉积前后的形貌特征.利用循环伏安技术研究了该银微粒电极对血红蛋白(Hb)的直接电化学行为.实验结果表明具有立体结构的银微粒对血红蛋白具有良好的电催化活性,实现了血红蛋白分子的直接电子转移.在1×10-6 mol·L-1~2×10-5 mol·L-1范围内,Hb氧化峰电流和Hb浓度有较好的线性关系.该银微粒电极制备简单,性能稳定,使用寿命相对较长,可有望用于蛋白质的分析测定和实现Hb在第三代生物传感器的开发和应用.     

血红蛋白在银微粒修饰镍铬电极上的直接电化学行为研究

以镍铬合金为基体电极,采用电沉积技术制备了银微粒电极.用扫描电镜表征了基体电极电沉积前后的形貌特征.利用循环伏安技术研究了该银微粒电极对血红蛋白(Hb)的直接电化学行为.实验结果表明具有立体结构的银微粒对血红蛋白具有良好的电催化活性,实现了血红蛋白分子的直接电子转移.在1×10-6 mol·L-1~2×10-5 mol·L-1范围内,Hb氧化峰电流和Hb浓度有较好的线性关系.该银微粒电极制备简单,性能稳定,使用寿命相对较长,可有望用于蛋白质的分析测定和实现Hb在第三代生物传感器的开发和应用.     

纳米介孔氧化硅对雌二醇的氧化增敏效应及其电化学检测研究

以十六烷基三甲基溴化铵(CTAB)为模板,正硅酸四乙酯(TEOS)为硅源,合成出纳米介孔二氧化硅(Me-so-SiO2),并制成修饰碳糊电极.用K3[Fe(CN)6]探针研究了修饰电极的传感特性,介孔二氧化硅不仅显著增加了碳糊电极的表面积,而且明显提高了其异相电子交换速率常数.研究了雌二醇的电化学行为,结果表明介孔二氧化硅对雌二醇的氧化有显著的增敏效应,明显地提高其氧化峰电流.考察了pH值、介孔二氧化硅用量、富集时间对雌二醇氧化信号的影响规律,建立了一种测定雌二醇的新电分析方法,其线性范围为1.0×10-7~7.0×10-6mol/L,检测限为6.0×10-8mol/L.将此方法用于注射液中雌二醇含量的测定,结果与标示量很好地吻合.     

介孔硅材料组装DNA电化学生物膜电极的制备

利用马来酰亚胺基类酯(Sulfo-SMCC)为异双功能偶联剂,将巯基修饰好的DNAzyme共价固定在已负载亚甲基蓝的氨基化介孔二氧化硅纳米材料上,并将壳聚糖混合后滴涂至碳糊电极表面上形成了电化学生物传感膜.利用循环伏安法、电化学阻抗法等对膜电极的制备过程进行了相应的表征.结果表明:基于巯基修饰的DNAzyme已成功固定于二氧化硅纳米材料上,并在碳糊电极表面上形成了稳定的膜.     

有序介孔分子筛的生物大分子固定化研究

利用介孔材料孔径和生物大分子的尺寸相当的特点,采用有序介孔分子筛SBA-15为载体实现对血红蛋白（Hb）和葡萄糖氧化酶（GOx）的固载,并考察其生物活性．紫外可见光谱和红外吸收光谱表明：以介孔分子筛SBA-15作为固定载体可以有效地吸附血红蛋白和葡萄糖氧化酶,且在吸附过程中很好的保持了蛋白质的天然二级结构．     

酪氨酸酶和漆酶的直接电化学研究

首次报道了吸附在活性炭表面的酪氨酸酶(Tyr)和漆酶(Lac)能进行准可逆的直接电化学反应，并保持对氧还原的电催化作用．本文所用的固定酶的方法具有简单、易于操作和酶活性保持良好等优点，可用于研究氧化还原蛋白质和酶的直接电化学和用于制备生物燃料电池的催化剂．     

Nanostructured ZnO for biosensing applications

Based on the unique properties, nanostructured ZnO could provide a stable immobilization for biomolecules retaining their biological activity. It has been recently developed as a nice candidate for the construction of biosensors with enhanced analytical performance. In this paper, we reviewed the progress in adapting nanostructured ZnO for several predominantly in biosensing applications based on enzymic reaction, immunoreaction, and molecular compitation. We also described several important considerations when working with nanostructured ZnO mainly centered on the fabrications of ZnO and appropriate strategies for biosensor construction (e.g. modified electrodes and multilayered immobilization).     

纳米银—金复合颗粒对葡萄糖生物传感器响应灵敏度的增强效应

以纳米银-金复合颗粒与聚乙烯醇缩丁醛(PVB)构成复合固酶膜基质,用溶胶-凝胶法固定葡萄糖氧化酶(GOD),组成葡萄糖生物传感器。实验表明,纳米颗粒可以使电极的响应电流从相应浓度的几十纳安增强到几千纳安。探讨了纳米颗粒效应在固定化酶中所起的作用,开辟了制备直接电子传递第三代生物传感器的新途径。     

Highly sensitive biosensors based on water-soluble conjugated polymers

Conjugated, conductive polymers are a kind of important organic macromolecules, which has found applications in a variety of areas. The application of conjugated polymers in developing fluorescent biosensors represents the merge of polymer sciences and biological sciences. Conjugated polymers are very good light harvesters as well as fluorescent polymers, and they are also “molecular wires”.Through elaborate designs, these important features, i.e.efficient light harvesting and electron/energy transfer, can be used as signal amplification in fluorescent biosensors. This might significantly improve the sensitivity of conjugated polymer-based biosensors. In this article, we reviewed the application of conjugated polymers, via either electron transfer or energy transfer, to detections of gene targets, antibodies or enzymes. We also reviewed recent efforts in conjugated polymer-based solid-state sensor designs as well as chip-based multiple target detection. Possible directions in this conjugated polymer-based biosensor area are also discussed.     

Synthesis and characterization of chiral mesoporous silica

Chirality is widely expressed in organic materials, perhaps most notably in biological molecules such as DNA, and in proteins, owing to the homochirality of their components (d-sugars and l-amino acids). But the occurrence of large-scale chiral pores in inorganic materials is rare. Although some progress has been made in strategies to synthesize helical and chiral zeolite-like materials, the synthesis of enantiomerically pure mesoporous materials is a challenge that remains unsolved. Here we report the surfactant-templated synthesis of ordered chiral mesoporous silica, together with a general approach for the structural analysis of chiral mesoporous crystals by electron microscopy. The material that we have synthesized has a twisted hexagonal rod-like morphology, with diameter 130-180 nm and length 1-6 micro m. Transmission electron microscopy combined with computer simulations confirm the presence of hexagonally ordered chiral channels of 2.2 nm diameter winding around the central axis of the rods. Our findings could lead to new uses for mesoporous silica and other chiral pore materials in, for example, catalysis and separation media, where both shape selectivity and enantioselectivity can be applied to the manufacturing of enantiomerically pure chemicals and pharmaceuticals.     

电流型酶电极生物传感器抗可逆抑制剂干扰的测定方法

电流型酶电极生物传感器在工业领域的应用中经常会遇到酶活性抑制剂干扰问题,在存在酶活性抑制剂的环境下,用酶电极生物传感检测,分析结果会显著偏低。根据电流型酶电极生物传感器的检测原理,设计了一种抗抑制剂干扰的测定方法,传感器正常定标完成后,先按正常方法进行测定,然后用样品加标样进行第二次测定,两次测定具有相同的酶抑制剂环境。通过两次测定结果比较,算出第二次测定所加标样的响应结果,计算出抑制剂对酶活的抑制系数,最终通过第一次测定结果和抑制系数计算出样品中待测物的浓度。这种测定方法能有效消除酶抑制剂对分析结果的影响,拓展酶电极生物传感器在工业领域中的应用范围。     

Visualizing single-molecule diffusion in mesoporous materials

Periodic mesoporous materials formed through the cooperative self-assembly of surfactants and framework building blocks can assume a variety of structures, and their widely tuneable properties make them attractive hosts for numerous applications. Because the molecular movement in the pore system is the most important and defining characteristic of porous materials, it is of interest to learn about this behaviour as a function of local structure. Generally, individual fluorescent dye molecules can be used as molecular beacons with which to explore the structure of--and the dynamics within--these porous hosts, and single-molecule fluorescence techniques provide detailed insights into the dynamics of various processes, ranging from biology to heterogeneous catalysis. However, optical microscopy methods cannot directly image the mesoporous structure of the host system accommodating the diffusing molecules, whereas transmission electron microscopy provides detailed images of the porous structure, but no dynamic information. It has therefore not been possible to 'see' how molecules diffuse in a real nanoscale pore structure. Here we present a combination of electron microscopic mapping and optical single-molecule tracking experiments to reveal how a single luminescent dye molecule travels through linear or strongly curved sections of a mesoporous channel system. In our approach we directly correlate porous structures detected by transmission electron microscopy with the diffusion dynamics of single molecules detected by optical microscopy. This opens up new ways of understanding the interactions of host and guest.     

Optimal design of label-free silicon“ lab on a chip” biosensors

This paper reported the optimal design of label-free silicon on insulator(SOI) "lab on a chip"biosensors.These devices are designed on the basis of the evanescent field detection principles and interferometer technologies.The well-established silicon device process technology can be applied to fabricate and test these biosensor devices.In addition,these devices can be monolithically integrated with CMOS electronics and microfuidics.For these biosensor devices,multi-mode interferometer(MMI) was employed to combine many stand-alone biosensors to form chip-level biosensor arrays,which enable realtime and label-free monitoring and parallel detection of various analytes in multiple test samples.This sensing and detection technology features the highest detection sensitivity,which can detect analytes at extremely low concentrations instantaneously.This research can lead to innovative commercial development of the new generation of high sensitivity biosensors for a wide range of applications in many fields,such as environmental monitoring,food security control,medical and biological applications.     

碳点的制备及其生物成像与生物检测应用

碳点(CDs)作为一种新的含碳荧光纳米粒子,由于其良好的生物相容性引起了广泛的关注。碳点不仅被用于生物成像探针,还被用来作为生物传感探针。目前,碳点的合成、结构与性质、荧光机理以及对生物相容性和生物应用的评价等方面都有了很大的进展。从碳点的合成方法、荧光性质以及在生物成像和生物检测应用方面进行了概述,重点阐述了碳点用于荧光比率传感的设计与构建,并对碳点研究的发展方向和应用前景进行了评述与展望。     

Direct imaging of the pores and cages of three-dimensional mesoporous materials

Mesostructured composite materials, with features ranging from 20 to 500 A in size, are obtained by the kinetically controlled competitive assembly of organic and inorganic species into nanostructured domains. Short-range order is limited, and long-range order is determined by weak forces such as van der Waals or hydrogen-bonding. Three-dimensional mesoporous materials obtained by removing the organic phase are of particular interest for applications such as catalysis and chemical sensing or separation, for which structural features such as cavity shape, connectivity and ordered bimodal porosity are critical. But atomic-scale structural characterization by the usual diffraction techniques is challenging for these partially ordered materials because of the difficulty in obtaining large (> 10 microm) single crystals, and because large repeat spacings cause diffraction intensities to fall off rapidly with scattering angle so that only limited small-angle data are available. Here we present a general approach for the direct determination of three-dimensional mesoporous structures by electron microscopy. The structure solutions are obtained uniquely without pre-assumed models or parametrization. We report high-resolution details of cage and pore structures of periodically ordered mesoporous materials, which reveal a highly ordered dual micro- and mesoscale pore structure.     

基于有机小分子近红外发光材料的研究进展

近红外有机发光二极管（Organic light emitting diodes, OLEDs）在夜间显示、生物传感和通讯等领域具有广阔的应用前景。其中有机小分子发光材料具有制作成本低、重现性好、结构和发光特性易调节、发光效率高等优点,根据其发光特性主要分为传统荧光材料、热激发延迟荧光（Thermally activated delayed fluorescence, TADF）材料、局部电荷转移杂化态（Hybrid locally-electron and charge-transfer, HLCT）材料等几类。本文主要综述了近五年来报道的基于有机小分子的近红外发光材料的研究进展,并根据不同的特性对材料进行了分类,对未来的发展前景进行了展望,为进一步合成各方面性能良好的近红外有机小分子发光材料提供了一定的理论基础。