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测定甲胎蛋白的非标记电位型免疫传感器 总被引:6,自引:0,他引:6
免疫分析是最重要的生物化学分析方法之一,是根据抗体与抗原或半抗原之间的专一性反应建立的分析方法,具有高度的选择性和极低的检测下限.电化学免疫传感器就是将免疫分析技术与电化学传感器相结合的一种新的免疫分析方法,具有免疫反应的高选择性和电化学分析的高灵敏性.甲脂蛋白(AFP)是人血清中含有的一种微量蛋白质,正常人血清中的含量在20ug/L以下,多种肝病都会引起甲胎蛋白含量的显著增加.甲脂蛋白能作为一种抗原与抗甲脂蛋白抗体反应,利用多克隆抗体免疫测定的甲胎蛋白总量,是临床化学检验上的一项重要指标.Aizawa等报… 相似文献
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研制了一种基于聚(3,4-乙烯二氧噻吩)(PEDOT)与天青Ⅰ(AzureⅠ)为基体的电化学免疫传感器,可灵敏检测甲胎蛋白(AFP)。在铂盘电极表面,电化学聚合PEDOT为基体,利用静电组装技术固定AzureⅠ和纳米金颗粒(nanoAus),将甲胎蛋白抗体(anti-AFP)组装到nanoAus的表面。采用辣根过氧化物酶(HRP)封闭非特异性吸附位点,制得电流型AFP免疫传感器。采用循环伏安、扫描电镜技术研究组装过程及电极性质,探讨了影响免疫传感器性能的因素。在优化实验条件下,电极响应与AFP的浓度在0.01~120μg/L的范围内呈线性关系,检出限为0.003μg/L。取临床血清样品用本方法检测AFP含量,得到的结果与临床常用的ELISA法得到的结果无显著性差异。 相似文献
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基于多层酶/纳米金固定甲胎蛋白免疫传感器的研究 总被引:21,自引:1,他引:21
>利用自组装技术和静电吸附作用, 将甲胎蛋白抗体(anti-AFP)固定在多层辣根过氧化物酶/纳米金及L-半胱胺酸修饰的金电极表面, 制备出用于检测甲胎蛋白抗原(AFP)的无试剂型免疫传感器. 通过交流阻抗技术、循环伏安法和计时电流法考察了电极的电化学特性, 并对该免疫传感器的作用机理及性能进行了详细的研究. 用计时电流法测得AFP的线性范围为1.0~10.0和10~200 ng•mL-1, 检出限为0.5 ng•mL-1. 实验结果表明, 该方法提高了抗体的固定量, 增强了传感器的灵敏度和稳定性, 且该传感器响应迅速、选择性好, 血清中常见抗原不干扰测定. 将其用于临床血清检验, 与放射免疫测定法(RIA)的符合率为86.7%. 相似文献
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报道了一种基于金纳米粒子(AuNPs)双重信号放大的高灵敏电化学免疫传感器,并应用于肝癌标志物甲胎蛋白(AFP)的检测。通过在丝网印刷电极(SPE)表面电沉积AuNPs提高电极的重现性,利用AuNPs的吸附作用固定AFP抗体,用于捕获样品中的待测AFP抗原,并进一步与固定了辣根过氧化物酶(HRP)标记检测抗体的纳米金免疫探针发生特异性结合,所形成的夹心免疫复合物可以催化底物得到响应电流。用扫描电镜(SEM)和微分脉冲伏安法(DPV)等技术研究电极组装过程以及电极的化学性质,讨论了影响免疫传感器性能的因素。在最优实验条件下,传感器的峰电流信号与AFP浓度在2.5~30ng/mL范围内呈良好的线性关系,检出限为0.16ng/mL。该传感器具有灵敏度高、成本低、仪器体积小的优点,具有较好的应用前景。 相似文献
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在金电极表面电沉积银为氧化还原探针,利用有机多孔材料(PTC-NH2)、纳米金(nano-Au)固载甲胎蛋白抗体(anti-AFP),制备出用于检测甲胎蛋白(AFP)的安培型免疫传感器。通过交流阻抗技术、循环伏安法研究了电极的电化学特性,考察了孵育时间、测试液pH值等实验条件对传感器性能的影响,并利用扫描电子显微镜(SEM)对电极的修饰过程进行了表征。该传感器对AFP有良好的电流响应,线性范围分别为1.0~20.0ng/mL和20.0~60.0 ng/mL,检测限为0.6 ng/mL。 相似文献
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建立了一种可同时检测双组分肿瘤标志物的免标记电化学免疫测定方法,检测了人血清样品中癌胚抗原(CEA)和甲胎蛋白(AFP)的含量. 在实验中,我们首先将二种电化学底物,二茂铁甲酸(FCA)和亚甲基蓝(MB),分别结合在离子液体(IL)修饰的介孔硅(MPS)的孔道内,制备FCA-IL-MPS和MB-IL-MPS复合材料;然后将免疫探针涂覆在掺铟氧化锡(ITO)电极表面的不同部分;最后将CEA单克隆抗体(anti-CEA)和AFP单克隆抗体(anti-AFP)分别负载到两种材料的孔内,制得CEA和AFP的免标记免疫探针(anti-CEA/FCA-IL-MPS,anti-AFP/MB-IL-MPS),组装得到双组分的免标记免疫传感器. 当该免疫传感器在含有抗原的样液中温育后,CEA和AFP抗原通过特异性免疫反应而结合到MPS孔道内. 由于形成的免疫复合物不导电且有一定的空间位阻,所以对电极表面的电子转移产生了阻碍. 根据ITO不同位置上响应电流的变化实现对CEA和AFP含量的同时测定. 本方法对CEA和AFP的检测线性范围分别为0.5~80 ng·mL-1和0.5~100 ng·mL-1,检测限分别为0.1 ng·mL-1和0.1 ng·mL-1(S/N=3). 离子液体具有高导电率,能实现所构建体系信号的放大,使制备的免疫传感器具有良好的灵敏度. 相似文献
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传染性病原体POCT对于及时有效控制传染病尤为关键。相比于传统检测方法,基于电化学免疫传感器的传染性病原体检测具有快速、灵敏、准确、易于小型化和集成化等优势,尤其适用于传染病POCT。新兴的纳米材料因其独特的理化性质可用于修饰传感器界面或作为生物分子的固载基质以及信号标记物等,有助于构建出高选择性和高灵敏度的电化学免疫传感器。在本文中,我们着重阐述了不同结构的纳米材料修饰的电化学免疫传感器在传染性病原体POCT检测中的应用,进一步介绍了基于纳米材料的电化学免疫传感器与不同检测技术联用在传染性病原体POCT中的应用,并对其发展前景做出了展望。 相似文献
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Okadaic acid (OA), a lipophilic phycotoxin is mainly produced by toxigenic dinoflagellates. The need to develop high performing methods for OA analysis able to improve the traditional ones is evident. In this work, a novel experimental methodology for label-free detection of OA was developed. Protein G magnetic beads (protein-G-MBs) modified gold electrode was used to immobilize anti-OA monoclonal antibody (anti-OA-MAb). Preliminary, colorimetric tests were performed in order to validate protein-G-MBs and anti-OA-MAb reaction. Electrochemical detection was carried out by differential pulse voltammetry in ferri/ferrocyanide solution. The limit of detection value obtained (0.5 μg L−1) validated the developed electrochemical immunosensor as a promising tool for routine use. The matrix effect and the recovery rate were also assessed with real samples, showing a good percentage of recovery. 相似文献
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《Analytical letters》2012,45(15):2615-2631
Abstract An amperometric immunoassay technique utilizing a highly dispersed immunoelectrode was developed. The assay method is based on the ‘sandwich’ scheme of immunointeraction. Rabbit IgG has been used as model analyte. Horseradish peroxidase labelled conjugates were employed coupled with amperometric detection of iodine reduction. Highly dispersed ULTI carbon serves as an immunosorbent and at the same time as an electrode material. It was found that the dependence of immunoelectrode response on the time of incubation during the second stage of ‘sandwich’ -based assay was complex. This is probably due to non-equilibrium immuno-agglomeration of sub-micron sized particles of immunosorbent. The improvement in the sensitivity of immunoelectrode by increasing the amount of immunosorbent was demonstrated. The increase in the magnitude of the immunoelectrode response is proportional to the increase of the amount of immunosorbent. A detection limit of 10 pM has been achieved with an overall assay time of less than 20 minutes. 相似文献
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Farré M Martínez E Ramón J Navarro A Radjenovic J Mauriz E Lechuga L Marco MP Barceló D 《Analytical and bioanalytical chemistry》2007,388(1):207-214
A new immunoassay for continuously monitoring atrazine in water has been developed. It uses a portable biosensor platform
based on surface plasmon resonance (SPR) technology. This immunoassay is based on the binding inhibition format with purified
polyclonal antibodies, with the analyte derivative covalently immobilized on a gold sensor surface. An alkanethiol self-assembled
monolayer (SAM) was formed on the gold-coated sensor surface in order to obtain a reusable sensing surface. The low detection
limit for the optimized assay, calculated as the concentration that produces a 10% decrease in the blank signal, is 20 ng/L.
A complete assay cycle, including regeneration, is accomplished in 25 min. Additionally, a study of the matrix effects of
different types of wastewater was performed. All measurements were carried out with the SPR sensor system (β-SPR) commercialised
by the company Sensia, S.L. (Spain). The small size and low response time of the β-SPR platform would allow it to be used
in real contaminated locations. The immunosensor was evaluated and validated by measuring the atrazine content of 26 natural
samples collected from Ebro River. Solid-phase extraction followed by gas chromatography coupled to mass spectrometric detection
(SPE–GC–MS) was used to validate the new immunoassay. 相似文献
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伏安免疫法检测牛奶中氯霉素残留 总被引:6,自引:0,他引:6
为探索用于现场检测牛乳中氯霉素(CAP)残留的高灵敏度及特异性强的免疫传感器方法,本实验在制备CAP单克隆抗体的基础上,以卵清蛋白-氯霉素(OVA-CAP)偶联物为包被抗原,并将其包被到聚苯乙烯反应板上;在孵育反应中,样品中的CAP与OVA-CAP竞争结合CAP单克隆抗体,洗涤后加入碱性磷酸酶(ALP)标记的二抗,经再次孵育及洗涤后加入对硝基苯磷酸(pNPP)底物液;反应终止后用线性导数伏安法记录pNPP水解产物的氧化峰电流。实验结果表明,用免疫传感法测试CAP的灵敏度高于传统的间接竞争酶联免疫吸附法(ELISA)。该方法检测CAP的检出限为0.064μg/L;检测线性范围为0.15~600μg/L,测试牛奶样品的平均回收率为89.8%。另外,由于免疫电化学传感器体积较小,便于携带,操作简单,可实现牛乳样品中CAP残留的现场检测。 相似文献
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压电免疫传感器用于C2型葡萄球菌肠毒素的测定 总被引:2,自引:0,他引:2
报道了用蛋白A-金电极法将抗体固化在3.58MHz和10HzAT切割的石英晶体上,制作的可重复使用的压电免疫传感器,测定C2型葡萄球菌肠毒素。最适PH值为7,并用不同的方法测试了晶体片上的抗体固化情况。 相似文献
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《Analytical letters》2012,45(14):2459-2474
Abstract An immunoassay technique based on a high surface area carbon immunoelectrode is described. Dispersed ULTI carbon serves as a carrier for immobilized antibodies and also as an electrode material. ‘Sandwich’ scheme of immunoassay of rabbit IgG (as a model analyte) has been used. Iodine formed as a result of the enzymatic oxidation of iodide by peroxidase-label has been detected amperometrically. Using dispersed carbon material as a solid support for immobilization of immunoagents and as an electrode material improves the efficiency of immuno-interaction and the sensitivity of electrochemical assay due to the high area-to-volume ratio of solid to liquid phases. The method based on ‘sandwich’ immunoassay is conducted at 21°C. The time of each stage of incubation (with analyte and with peroxidase-antibody conjugate) is 10 min. Electrochemical detection of peroxidase-labeled immuno-complex does not exceed several minutes. The technique allows fast determination of rabbit IgG with a low detection limit, in the nanomolar range. By comparison, the conventional technique based on ELISA, being performed under the same conditions (temperature and incubation time) does not allow any quantitative determination of IgG for concentration lower then 100 nM. 相似文献
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以电化学聚合法在石墨电极上获得聚间苯二胺膜(PMPD),并建立了快速、灵敏的膜质量检验方法:辣根过氧化物酶(HRP)标记PMPD膜-邻苯二胺(OPD)反应法,对聚合、活化等条件作了研究;进一步制成了检测乙肝表面抗原(HBsAg)和检测鼠免疫蛋白(MIgG)的免疫电极,对血清样品作了检测.结果表明:2.5V电池电压下,于含0.12mol/LMPD的1.2mol/LH2SO4中聚合20min,再以30g/L浓度的戊二醛活化4h,所得HRP标记电极及免疫电极重现性好;检测HB-sAg和MIgG的线性范围分别为0.1~3.2mg/L及0.1~10mg/L 相似文献