生物小分子巯基化合物荧光探针研究进展

细胞内的小分子巯基化合物在诸多生理过程中扮演重要角色.分子荧光探针具有灵敏度高、选择性好、生物相容性好、实时原位监测等优点.因此,构建可以选择性检测巯基化合物的荧光探针具有重要的生物学和医学意义.根据荧光探针与巯基化合物的反应类型总结了近几年来小分子巯基化合物荧光探针的设计策略和研究进展.     

快速检测巯基化合物的新方法

本文利用巯基化合物对异硫氰酸荧光素(FITC)的荧光猝灭作用,建立了一种快速检测巯基化合物的新方法。实验中发现,巯基化合物能够猝灭FITC的荧光,据此发展了检测巯基化合物的新方法。本方法的用时非常短,在优化了pH、反应时间等条件后能在5min之内完成对巯基化合物的检测。在最佳条件下对实际样品的测试取得了很好的效果,且具有良好的选择性。对谷胱甘肽(GSH)、高半胱氨酸(Hcy)、半胱氨酸(Cys)、二硫苏糖醇(DTT)的检测限分别达到3.08、2.66、1.28和1.92μmol/L。     

巯基化合物自组装单分子层的研究进展

超薄层有机材料的制备与研究￣［１］受到人们的广泛关注，有关研究结果和进展正在不断丰富着人类对表面结构与性质关系的理解和认识。与其它制备单分子层的技术相比，自组装技术是一种较新的制备方法，在８０年代后期，迅速成为有关学科的研究焦点。采用此技术制备的超薄层体系主要有两类：一类是巯基化合物在金、银、铜、铂表面吸附形成的单层￣［２，３］，一类是在硅、玻璃、金属氧化物表面通过硅烷化反应形成的单层￣［４］。其中，硫醇类分子自组装单层（ｓｅｌｆ－ａｓｓｅｍｂｌｅｄｍｏｎｏｌａｙｅｒ，３ＡＭ）是被研究得最为广泛和深入的体系。本文主要综述十年来巯基化合物ＳＡＭ的研究进展。     

巯基化合物分离富集技术的应用进展

根据负载(或鳌合)巯基的物质不同,分别对巯基棉、巯基葡聚糖凝胶、巯基纸、巯基活性炭和巯基树脂等分离富集剂的制备、应用及机理进行了综述。     

基于不同电化学探针检测乳腺癌基因片段电化学传感器的研究

构建了以3种不同电活性物质(铁氰化钾平衡电对、亚甲基蓝和六氨合钌)为电化学信号探针,检测乳腺癌基因片段(乳腺癌DNA)的电化学传感器。利用吸附作用将探针ss DNA固定于金纳米-多壁碳纳米管-Nafion复合纳米材料修饰金电极表面,制备了DNA电化学传感器。采用循环伏安法、电化学阻抗法和微分脉冲伏安法,对DNA电化学传感器进行表征和定量分析。实验结果表明,在5 mmol/L K3[Fe(CN)6]-5mmol/L K4[Fe(CN)6]平衡电对电化学探针检测液中,乳腺癌DNA的线性范围为0.1~500.0 nmol/L,其检出限(S/N=3)为0.03 nmol/L。以20μmol/L亚甲基蓝为电化学探针检测液时,乳腺癌DNA的线性范围为1.0~500.0 nmol/L,检出限为0.3 nmol/L。利用50μmol/L六氨合钌电化学探针检测时,乳腺癌DNA的线性范围为1.0~500.0 nmol/L,检出限为0.3 nmol/L。3种电化学探针中,利用铁氰化钾平衡电对探针检测乳腺癌DNA的检出限最低,线性范围最宽。该传感器可用于其他DNA的检测分析。     

巯基化合物的电位分析法研究

本文对用直接电位法测试巯基化合物的可行性进行了研究。发现由硫化银，硫化汞等金属难溶盐制成的膜电极对６－巯基嘌呤，高胱氨酸，１，４－二巯基苏糖经合物可产生响应，其斜率为３０－１１０ｍＶ／ｐＣ，电极响应时间，则为十几秒至十几分钏，测试结果具有较好的重现性。因此，该方法可望得到应用。文中还对有关响应机理进行了探讨。     

电化学和石英晶体微天平研究2-巯基苯并咪唑的电化学氧化反应

应用循环伏安研究水相中２－巯基苯并咪唑的电化学氧化过程,考察了ｐＨ值、浓度等的影响,并应用电化学石英晶体天平（ＥＱＣＭ）监测了此氧化成膜的过程。结果表明此反应为一电子过程。结合表面光电子能谱（ＸＰＳ）对此氧化膜进行了初步表征。     

自催化型石墨烯-金纳米探针的制备及用于非小细胞肺癌CYFRA21-1电化学检测

早期诊断对提高肺癌患者生活质量和延长生存期至关重要。本研究通过热解柠檬酸与谷氨酰胺的混合物得到谷氨酰胺功能化石墨烯量子点(Gln-GQD),所制备的Gln-GQD与HAuCI₄反应形成Gln-GQD/Au复合物。Gln-GQD作为还原剂将Au³⁺转化为Au⁰,最终形成Au纳米晶体。Gln-GQD还作为稳定剂被固定于Au纳米晶体的表面,从而实现Gln-GQD与Au的杂交。扫描电镜、透射电镜、X-射线衍射和红外光谱分析表明,Gln-GQD/Au平均粒径为(31.2±0.15) nm,具有立方金的晶体结构,Au和N元素均匀分布于球形粒子表面,含有丰富的OH、NH、COOH等功能基团。将发夹DNA 2 (H2)通过AuS键连接到Au纳米粒子表面,再利用EDC/NHS活化实现Gln-GQD的-COOH与硫堇(Thi)的氨基缩合,得到H2-Gln-GQD/Au-Thi氧化还原探针,基于此构建了无酶放大电化学传感平台。在CYFRA21-1存在下,此传感平台的H2可与预先修饰在金电极表面的发夹DNA 1(H1)杂交,释放出一个CYFRA...     

基于纳米探针的肿瘤细胞电化学传感研究进展

纳米材料在纳米尺度下具有独特的光、电、磁等性质,已广泛应用于细胞电化学传感的研究,该研究工作提高了肿瘤细胞检测的灵敏度与特异性,为癌症的早期诊断与治疗提供了新的方法与思路。本文综述了基于纳米探针进行肿瘤细胞电化学传感研究的工作,总结了纳米电化学细胞传感技术在肿瘤细胞固定、识别、检测及诱导凋亡等方面的研究进展,并对未来研究动向进行了展望。     

HCl溶液中巯基杂环化合物对铜的缓蚀作用

铜缓蚀剂;HCl溶液中巯基杂环化合物对铜的缓蚀作用;极化曲线;电化学阻抗;计算     

Carbon Nanotube‐Adsorbed Electrospun Nanofibrous Membranes as Coating for Electrochemical Sensors for Sulfhydryl Compounds

A new method to modify electrodes with carbon nanotubes (CNT) was developed. Multiwalled carbon nanotubes (MWNT) were adsorbed on the electrospun nylon‐6 nanofibrous membranes (Ny‐6‐NFM) and used as a coating to modify conventional glassy carbon electrodes. The modified electrode was designed for the amperometric detection of sulfhydryl compounds with the potential held at +0.3 V vs. Ag/AgCl. The modified electrode showed a linear response for cysteine up to 0.4 mM (R²=0.997), with a sensitivity of 5.1 µA/mM and a detection limit of 15 µM. Other sulfhydryl compounds showed similar results. After use, the Ny‐6‐NFM was easily peeled off, leaving the bare electrode surface back to its original electrochemical behaviour. This is the first attempt to use a disposable membrane functionalized with MWNT for electroanalytical purposes.     

基于阴离子-石墨嵌层化合物的电化学电容器

石墨可以在高电势下电化学可逆存储阴离子,有望在高电压储能器件中担当正极材料.本文介绍了基于阴离子-石墨嵌层化合物型正极材料的高比能电容器的研究进展,剖析了影响电容器性能的各方面因素,探讨了一系列表征相关电极材料储能机制的方法和手段,揭示了溶剂化效应对阴离子插嵌石墨正极电化学行为的关键性作用.并进一步概述了该种正极材料近年来在新型储能器件-双离子电池中的发展态势,展望了其应用前景和即将面临的潜在问题.     

基于吐温-姜黄素复合物的荧光探针检测脂肪酶活性

本实验以姜黄素作为信号探针,吐温作为脂肪酶的底物和包裹姜黄素的载体,建立了一种检测脂肪酶活性的荧光法.实验发现,采用0.3 mmol/L吐温40,25 μmol/L姜黄素,并且脂肪酶水解吐温40的时间为35 min时,姜黄素的荧光强度变化值与脂肪酶浓度在0.002 ～0.05 mg/mL和0.05～0.25 mg/mL范围内呈现线性关系,检出限为0.6 mg/L (S/N=3).此探针在高通量检测脂肪酶活性以及与脂肪酶相关疾病检测领域中有较好的应用前景.     

Voltammetric Detection of Free Sulfhydryl Compounds in Food Flours

Cyclic voltammetry has been applied to the rapid measurement of free SH‐compounds in food flours samples. The protocol is based on the electrochemical adaptation of the Ellman's test, where the DTNB reacts with SH‐compounds present in food flour extracts, resulting in the equimolar production of the electroactive compound TNBA which, in turns, reacts with phenylendiamine. The reaction is found to provide an analytical signal from which to quantify indirectly free sulfhydryl compounds in flours extracts.     

Detection of the Sulfhydryl Group in Proteins by Raman Scattering Spectroscopic Method

Detection of the sulfhydryl group in proteins has previously been difficult. However, the group can be detected quite easily by Raman spectroscopy due to its distinct vibrational band at 2560-2580 cm^?1 region. This region is free of interference from other vibrational bands of protein. Detection of SH in tobacco mosaic virus coat protein and two sea snake neurotoxins was made to illustrate the application of Raman spectroscopy.     

An Electrochemical Immunobiosensor for Direct Detection of Veterinary Drug Residues in Undiluted Complex Matrices

An electrochemical immunobiosensor is developed that allows the detection of small molecules, such as drugs, in undiluted complex samples, with no washing or rinsing steps via a displacement assay. This is achieved using an interface comprised of a mixed layer of oligo(phenylethynylene) molecular wire (MW), to allow electrochemical communication, and oligo(ethylene glycol) (OEG) to control the interaction of proteins and electroactive interferences with the electrode surface. The mixed layer is formed from in situ‐generated aryl diazonium cations. To the distal end of the MW, a redox probe 1,1′‐di(aminomethyl)ferrocene (FDMA) is attached followed by an epitope (the structural feature the antibody selectively recognizes) to which an antibody would bind. Association or dissociation of the antibody with the sensing interface causes a modulation of the ferrocene electrochemistry. Antibody complexed electrodes are exposed to samples containing spiked enrofloxacin (unbound target analyte), in milk and environmental water and interrogated using square wave voltammetry (SWV). The lowest detected concentration of free enrofloxacin was 10 pg mL^?1 in phosphate buffer, 50 mM, pH 7. For free enrofloxacin detection in undiluted complex matrices, by adding disodium EDTA (50 mM), the recovery obtained was 94.1 % in skim milk and 88.5 % in stream water, respectively as compared to clean phosphate buffer. The immunobiosensor response time was 10–15 minutes. The sensor performance in milk was shown to be superior to a standard method based on Liquid Chromatography Mass Spectroscopy (LC‐MS/MS).     

基于萘酰亚胺的生物硫醇探针的合成及对含硫醇氨基酸的检测

合成了以4-羟基萘酰亚胺为荧光团,2,4-二硝基苯磺酰氧基为特异性识别基团的生物硫醇探针4-(2,4-二硝基苯磺酰氧基)-正丁基-1,8-萘酰亚胺(DNSBN).吸收光谱和荧光光谱结果表明, DNSBN对半胱氨酸(Cys)、同型半胱氨酸(Hcy)和谷胱甘肽(GSH)3种生物硫醇分子具有高效的检测识别能力,不受其它17种天然氨基酸的干扰.同时,通过荧光滴定实验证实了此探针是一种比率型探针,555 nm处的荧光强度与溶液中的生物硫醇分子浓度在0 ~ 20 μmol/L范围内呈良好的线性关系,对Cys、Hcy和GSH的检出限(3σ)分别为25.9、92.0和77.9 nmol/L.而吸收光谱、荧光光谱和质谱表征数据显示,生物硫醇与2,4-二硝基苯磺酸酯发生亲核取代反应并导致磺酸酯的分解.随着识别基团的解离,探针分子的d-PeT (donor-excited photoinduced electron transfer) 效应被解除,并出现非常明显的比色与荧光变化.HeLa细胞成像实验表明,探针DNSBN具有良好的生物相容性,能够对细胞外源性生物硫醇分子进行检测.     

Mycotoxins Detection Based on Electrochemical Approaches

Electrochemical biosensors have attracted much attention in mycotoxin bioanalysis. In this review, three electrochemical biosensor technologies for mycotoxins were reviewed, including general electrochemistry, photoelectrochemistry, and electrochemiluminescence. Based on the classification of multiple electrochemical detection methods, the design schemes, recognition mechanism and probe materials were described in detail. Moreover, the characteristics and limitations of these electrochemical biosensors were summarized. The challenges and future trends of electrochemical biosensor development in mycotoxin bioanalysis were also briefly discussed in the end. This review is expected to provide some inspirations for point-of-care testing in electrochemical sensors for mycotoxins and further electrochemical analysis application.     

Electrochemical Detection of Avian Influenza Virus Genotype Using Amino‐ssDNA Probe Modified Gold Electrodes

A DNA biosensor for the detection of specific oligonucleotide sequences of Avian Influenza Virus type H5N1 has been proposed. The NH₂‐ssDNA probe was deposited onto a gold electrode surface to form an amide bond between the carboxyl group of thioacid and the amino group from ssDNA probe. The signals generated as a result of hybridization were registered in square wave voltammetry and electrochemical impedance spectroscopy in the presence of [Fe(CN)₆]^3?/4? as a redox marker. The genosensor is capable to determine 20‐mer and 180‐bp (PCR products) oligonucleotides complementary sequences with detection limit in the fM range. The genosensor displays good selectivity and sensitivity. The 20‐mer as well as 180‐bp oligonucleotides without a complementary sequence generate very low signal.     

有机杂环化合物在金属表面的化学及电化学聚合

介绍了2004年国家自然科学奖二等奖获奖项目“有机杂环化合物在金属表面的化学及电化学聚合”的主要科学内容、学术界的引用评述以及在学科发展、人才培养方面的成果。