表面等离子体共振生物传感器连续检测莱克多巴胺

利用表面等离子体共振生物传感器对莱克多巴胺抗体与固定在芯片表面的莱克多巴胺衍生物的相互作用进行了分析,解离常数为2.56×10-6s-1。根据一定范围内相对响应值和时间近似呈线性关系的动力学特性,建立了连续检测的方法,从而简化了实验步骤,有利于提高芯片的使用寿命。检测莱克多巴胺采用抑制法,将莱克多巴胺衍生物固定在芯片的表面,莱克多巴胺抗体与样品混合后流过芯片的表面,所得相对响应值与样品中莱克多巴胺的浓度成反比。单个样品的检测时间设定为15min,对应的检出限小于4μg/L。     

表面等离子体共振法检测人血清白蛋白抗体活性

表面等离子体共振法是研究生物大分子间相互作用的有效方法之一,和非直接方法（如酶联免疫）相比,具有实时、快速和免标记等特点。我们在甲羧基化葡聚糖修饰的传感片表面直接交联固一人血清白蛋白（ＨＳＡ）,用于ａｎｔｉ－ＨＳＡ抗体活性的检测,并用Ｈ３ＰＯ４（０．１ｍｏｌ／Ｌ）溶液再生。结果表明表面等离子体共振（ＳＰＲ）生物传感器能快速实时检测ａｎｔｉ－ＨＳＡ抗体的活性,且传感片能够重复使用１００次以上。     

表面等离子体共振生物传感器及其应用

表面等离子体共振(Surface Plasmon Resonance,SPR)生物传感器在检测生物分子特异性结合方面具有的高灵敏度、免标记及检测快速等优点,使其在过去的20年中在生命科学、医药科学、食品安全等领域取得了快速发展。本文对SPR生物传感器进行了简要介绍,并着重对其在生命科学、医药学、环境分析、食品安全等领域的应用进行了综述,最后对该领域的研究前景进行了展望。     

纳米金颗粒增强信号的表面等离子体共振生物传感器用于甲氧檗因高灵敏检测的研究

报道了一种基于表面等离子体共振(SPR)生物传感器的高灵敏检测抗癌药物甲氧檗因的新方法. 分别在纳米金颗粒和金膜表面修饰富含腺嘌呤(A)的DNA链, 当存在甲氧檗因时, 由于一个甲氧檗因分子可与4个A碱基相结合, 从而使得修饰在纳米金颗粒和金膜表面的DNA形成稳定的双链结构, 进而将功能化纳米金颗粒捕获在金膜表面. 由于纳米金颗粒与金膜之间的电场耦合效应可增强SPR信号, 从而可实现对小分子甲氧檗因的高灵敏、特异性检测. 本方法的检测下限低至0.07 pmol/L, 相对比色法和荧光法而言, 降低了约5～6个数量级. 以4种药物(盐酸小檗碱、青霉素G、硫酸庆大霉素、5-氟尿嘧啶)作为对照考察了该传感器的选择性, 结果表明该传感器具有较好的选择性.     

应用表面等离子体共振技术在传质控制下检测铁蛋白

应用表面等离子体共振技术在传质控制下对铁蛋白进行了检测。铁蛋白的分子量较高,传质较慢;通过增大抗体固定量和控制流速,使抗原-抗体间的反应在质量传输控制下进行。利用结合速率分析法对不同浓度的铁蛋白进行了灵敏检测,其线性范围为20~800μg/L,显著宽于“三明治”分析法。本方法检出限为20μg/L。用pH 2.0的甘氨酸-盐酸缓冲液再生,使同一传感片可重复使用50次以上。     

表面等离子共振技术测定生长抑素含量

应用表面等离子共振(SPR)技术建立了一种快速检测生长抑素(SST)含量的方法, 在pH=5.0, SST浓度为75 μg/mL的最佳偶联条件下, SST在CM5芯片上的偶联值为1231.7 Response unit(RU). 选择117.7 nmol/L作为固定的大分子量抗体浓度, 用抗原-抗体-抗原结合法检测SST纯品, 在20~2000 pg范围内, SST纯品含量和RU值之间可建立直线相关的标准曲线, R=-0.824, p<0.05, 变异系数CV=1.3%. 所建立的方法测定SST含量的范围为20~2000 pg, 重复性好且可随时检测, 有望用于SST的临床即时检测.     

Detection of Penicillin via Surface Plasmon Resonance Biosensor

A method of using Au colloid to capture the decomposed product of penicillin, penicillamine, on a surface plasmon resonance(SPR) biosensor for the quantitative determination of penicillin was developed. Based on the decomposition of penicillin to generate penicillamine and penilloaldehyde, a high sensitive biosensor for detecting penicillin was also developed. In our experiment, it was penicillamine rather than penicillin that has been measured. This is because penicillamine contains a functional group that makes it self-assembling on Au colloid to increase the molecular weight so as to improve the surface plasmon resonance signal. On a UV-Vis spectrophotometer, a high concentration of penicilliamine-Au complex was determined, indicating that penicillamine was already well combined with Au colloid. The method, using the combination of Au colloid with penicillamine, proved to detect penicillin.     

Imaging Array SPR Biosensor Immunoassays for Sulfamethoxazole and Sulfamethazine

A homemade array surface plasmon resonance (SPR)-based imaging biosensor was used to develop sensitive and fast immunoassays to determine sulfamethoxazole (SMOZ) and sulfamethazine (SMT) in buffer. Two conjugations of sulfonamide-bovine serum albumin (BSA) were separately immobilized on two different rows of the array chip with one row as reference. The immobilization was carried out in the instrument to monitor the quantity of the conjugations immobilized. The antibody mixed with the sulfonamide in the buffer was injected over the surface of the chip to get a relative response which was inversely proportional to the concentration of the sulfonamide in the PBS buffer. Two calibration curves were constructed and the limit of detection for sufamethoxazole in buffer was 3.5 ng/mL and for sulfamethazine 0.6 ng/mL. The stability and specificity of the antibody were also studied. The monoclonal antibody did not bind with BSA.     

Indirect Inhibitive Immunoassay for Estradiol Using Surface Plasmon Resonance Coupled to Online In-Tube SPME

Abstract

A new immunoassay method using surface plasmon resonance (SPR) coupled with online, in-tube, solid-phase microextraction (SPME) was developed and applied to detect estradiol in human serum and seawater using an indirect inhibitive immunoassay format. The binding of antibody was inhibited by estradiol in a dose-dependent manner, and the calibration curve was generated by linear fit over the range of 0.3125–20.0 ng/ml, with a correlation coefficient of R² = 0.99996. The detection limit (S/N = 3) was 0.17 ng/ml. This study demonstrates, for the first time, the feasibility of in-tube SPME-SPR to determine estradiol in human serum and seawater.     

利用表面等离子体谐振生物传感器实时监测酶促反应的新方法

在理论上，基于表面等离子体谐振(surface plasmon resonance，SPR)生物传感器的酶促反应检测分析方法被证明是可行的。这种方法与通常SPR传感器检测技术不同，它消除通常当成检测目标的表面吸附信号，而检测通常认为是噪声的本体折射率变化信号，以此实时检测酶促反应的动态过程。利用自产的SPR2000生化分析仪，在检测胶原酶I降解胶原I的实验中检测到了反应引起的反应液本体折射率的上升变化，从而证明了这种新颖的检测方法实际上也是可行的。这种技术一旦成熟，可以用来进行酶学分析及酶靶标的药物筛选研究开发。     

表面等离子体子共振传感器研究谷胱甘肽过氧化物酶模拟物与底物的结合作用

采用表面等离子体子共振(SPR)传感装置,固定入射角,以波长为变量,以CCD为检测系统,用对金有较强吸附作用的谷胱甘肽(GSH)为基底膜,研究了GSH分别与谷胱甘肽硫转移酶(GST)与小分子GPX模拟酶2-位-碲桥联-β-环糊精(2-TeCD)的动力学过程,计算得GSH与GST的结合常数为2.82×106L/mol;GSH与2-TeCD的结合常数为3.92×102L/mol。结果表明:本方法可以用来评价小分子GPX模拟酶与底物的结合程度,简便快捷,不需标记样品,可以实时监测反应过程,作用后酶与底物易于分离,分离后的酶可以继续使用等优点。     

Development of a Surface Plasmon Resonance-Based DNA Sensor and Its Application for Screening DNA-Targeted Anticancer Drugs

In this paper, we present a surface plasmon resonance (SPR)-based sensor for measuring DNA hybridization and DNA/small-molecule interactions. A mixed self-assembled monolayer (SAM) was used to optimize the biosensor sensitivity. It was observed that the mixed SAM formed by mixing 10 mM of 16-mercapto-1-hexadecanoic acid (16-MHA) and 6-mercapto-1-hexanol (6-MCH) at a 1:10 molar ratio showed the best results. Subsequently, avidin was attached to the carboxyl groups on the SAM to serve as a binding element for biotinylated single-stranded (ss)DNA. The ssDNA-coated sensor was first evaluated as a nucleic acid biosensor through a DNA-DNA hybridization assay for synthetic 28-mer ssDNA. A linear calibration curve was observed in the range of 0.25–2.5 µg/mL. Non-complementary DNA induced no significant SPR angle shift, which demonstrated the specificity of the assay. Secondly, the sensor was used to monitor the binding kinetics of DNA/small-molecule interactions in real time. The dissociation constant between immobilized DNA and sanguinarine was determined to be 8.0 × 10^?6 M. This complies with most data from the literature. In addition, the sensor could be regenerated with 0.01 M HCl and would be feasible for multiple testing. In conclusion, the experimental approach described in this study allows analysis of molecular interactions between DNA-binding drugs and selected targeted DNA sequences.     

表面等离子体共振免疫传感器在蛋白质检测中的应用及其研究进展

表面等离子体共振(SPR)技术是20世纪90年代发展起来的一种新型技术,应用SPR原理可检测生物传感芯片上配位体与分析物之间的相互作用情况,在生命科学、医疗检测、药物筛选、食品检测及环境监测等领域具有广泛的应用需求.SPR技术可与免疫传感器结合,利用抗原抗体的特异性反应可用于各种蛋白质抗原的检测.本文重点总结了SPR免疫传感器在食品及医疗领域蛋白质检测的应用,综述了近年来SPR免疫传感技术在这该领域的研究热点及进展.     

用表面等离子体子共振传感器测定人心肌肌钙蛋白I的研究

采用自组装表面等离子体子共振(SPR)传感装置,固定入射角,以波长为变量,以CCD为检测系统,用对金和抗体均有较强吸附作用的葡萄球菌A蛋白作为基底膜,观测了人心肌肌钙蛋白I的抗体和抗原之间免疫反应的动力学过程,并进行了人心肌肌钙蛋白I的定量测定.结果表明,人心肌肌钙蛋白I的浓度在5.0～50μg/L范围内与传感器的响应值呈线性关系.     

贵金属纳米材料的液相合成及其表面等离子体共振性质应用

金属中的表面等离子体共振是描述其导带电子在电磁场作用下集体振荡的一个物理概念。金属纳米颗粒由于其表面等离子体共振性质表现出独特的光学应用。本文在相关研究的基础上,综述了具备表面等离子体共振性质的不同形貌及多种复合结构的贵金属纳米材料的制备和应用,并对其发展及应用前景进行了展望。     

一种糖生物传感芯片制备的新方法

建立了一种用于SPR生物传感器分析的糖生物传感芯片制备新方法。在已二胺大量过量的情况下，海洋硫酸多糖911还原末端的半缩醛基与已二胺的一个氨基进行还原胺化反应，引入一个伯氨基，该伯氨基进一步与sulfo-NHS-biotin反应，使911的还原末端生物素化，通过预偶联到GM5芯片上的链酶抗生素抗体，以俘获法将生物素化的911固定到芯片表面。该方法避免已有固定方法对糖内部结构的破坏，减少了非特异性耦联，能更好地反映糖类与其它分子的相互作用特性，且操作简单，适应于多数具有还原末端的糖类。911经该方法固定后，利用SPR生物传感器，研究了其与HIV gp120蛋白V3区的相互作用动力学，初步证明两者之间存在强烈的相互作用，KA与KD分别为：2.25e5与4.44e-6。     

贵金属纳米材料的液相合成及其表面等离子体共振性质应用

金属中的表面等离子体共振是描述其导带电子在电磁场作用下集体振荡的一个物理概念。金属纳米颗粒由于其表面等离子体共振性质表现出独特的光学应用。本文在相关研究的基础上,综述了具备表面等离子体共振性质的不同形貌及多种复合结构的贵金属纳米材料的制备和应用,并对其发展及应用前景进行了展望。     

Immunosensor Based on Surface Plasmon Resonance for Antigen Recognition

A novel immunosensor based on surface plasmon resonance（SPR） has been developed for the recognition of antigen. The sensor was designed on the basis of the fixed angle of incidence and measuring the reflected intensities in a wavelength range of 430--750 nm in real-time. An ultra-bright white light-emitting diode（LED） was used as the light source. Molecular self-assembling in solution was used to form the sensing membrane on gold substrate. It has been seen that the sensitivity of the SPR sensor with 3-mercaptopropionic acid（MPA）/protein A（SPA） sensing membrane is considerably higher than that with MPA or SPA modified sensing membrane. The kinetic processes on the sensing membrane were studied. The human B factor（Bf）, an activator of complement 3（C3）, was recognized among the other antigens. This sensor can also be used for other antigen/antibody or adaptor/receptor recognition. Under optimized experimental conditions, the sensor has good selectivity, repeatability, and reversibility.     

Studies on Interactions of Antibiotics with Serum Albumin by Surface Plasmon Resonance Biosensor

Introduction Humanserumalbumin(HSA)isawell known transportproteinforavarietyofmoleculesandions[1].Thebindingofadrugtoserumalbuminhasimportant pharmacokineticconsequencesbecauseitinfluences distribution,excretionandpharmacologicaleffectsof thedruginthebody…     

表面等离子体子共振生物传感器用于乙肝表面抗原的测定

运用自行研制的表面等离子体子共振（SPR）生物传感器，采用自组装成膜技 术并以戊二醛作偶联剂，在传感片表面修饰HBsAg单克隆抗体，将其用于乙肝表面 抗原（HBsAg)的检测。实验结果表明SPR生物传感器对HBsAg的检出限为0.06ng/mL 。与传统的酶联免疫吸附试验（ELISA）相比，SPR生物传感器的检出灵敏度明显高 于ELISA法。用该SPR生物传感器对HBsAg质控血清与纯化的HBsAg溶液进行比较检测 ，结果表明该SPR生物传感器对HBsAg具有好的特异选择性。