Optical resonance-enhanced absorption-based near-field immunochip biosensor for allergen detection
Optical resonance-enhanced absorption-based near-field immunochip biosensor for allergen detection
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DOI：
10.1021/ac702107k
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发表时间：
2008-04-15
影响因子：
7.4
通讯作者：
Pittner, Fritz
中科院分区：
文献类型：
--
作者：
Maier, Irene;Morgan, Michael R. A.;Pittner, Fritz
建立了一种快速检测复杂食品基质中过敏原的光学免疫芯片生物传感器，并对鸡蛋白色、卵清蛋白和卵类粘蛋白过敏原的检测进行了评价。作为传感器设计基本原理基础的光学近场现象被称为共振增强吸收（REA），其利用金纳米颗粒（Au NPs）作为高灵敏度干涉设置中的信号换能器。使用这种方法，一种新的，简单的，快速的比色固相免疫分析的平面芯片基板上实现了直接和夹心测定格式，与检测系统，不需要任何仪器读出。半定量免疫化学反应是分析员肉眼直接可见的。该生物传感器显示浓度依赖性显色通过捕获抗体功能化的Au纳米粒子上的过敏原包被的芯片，并具有1 ng/mL的检测限。为了建立一个快速的方法，我们利用的物理化学微环境的Au NP-抗体生物缀合物被绑定直接在一个相互作用的聚（苯乙烯-甲基丙烯酸甲酯）夹层由固定化抗原。在直接测定形式中，在“软”非变性条件下，变应原的包被时间仅为5 min，就足以获得准确的再现性和灵敏度。总之，基于REA的免疫芯片传感器易于制造，其性能具有可重复性和选择性，具有最低的技术要求，并且将能够在技术和医学应用中高通量筛选亲和结合相互作用。
An optical immunochip biosensor has been developed as a rapid method for allergen detection in complex food matrixes, and its application evaluated for the detection of the egg white allergens, ovalbumin and ovomucoid. The optical near-field phenomenon underlying the basic principle of the sensor design is called resonance-enhanced absorption (REA), which utilizes gold nanoparticles (Au NPs) as signal transducers in a highly sensitive interferometric setup. Using this approach, a novel, simple, and rapid colorimetric solid-phase immunoassay on a planar chip substrate was realized in direct and sandwich assay formats, with a detection system that does not require any instrumentation for readout. Semiquantitative immunochemical responses are directly visible to the naked eye of the analyst. The biosensor shows concentration-dependent color development by capturing antibody-functionalized Au NPs on allergen-coated chips and has a detection limit of 1 ng/mL. To establish a rapid method, we took advantage of the physicochemical microenvironment of the Au NP-antibody bioconjugate to be bound directly over an interacting poly(styrene-methyl methacrylate) interlayer by an immobilized antigen. In the direct assay format, a coating time with allergen of only 5 min under "soft" nondenaturing conditions was sufficient for accurate reproducibility and sensitivity. In conclusion, the REA-based immunochip sensor is easy to fabricate, is reproducible and selective in its performance, has minimal technical requirements, and will enable high-throughput screening of affinity binding interactions in technological and medical applications.