Surface chemical approach to single-step measurement of antibody in human serum using localized surface plasmon resonance biosensor on microtiter plate system.
Surface chemical approach to single-step measurement of antibody in human serum using localized surface plasmon resonance biosensor on microtiter plate system.
复制标题
使用微量滴定板系统上的局部表面等离子共振生物传感器单步测量人血清中抗体的表面化学方法。
DOI：
10.1007/s00216-014-7833-5
复制
发表时间：
2014
影响因子：
4.3
通讯作者：
Maekawa T.
中科院分区：
文献类型：
--
作者：
Yamamichi J;Ojima T;Iida M;Yurugi K;Imamura T;Ashihara E;Kimura S;Maekawa T.
在临床环境中，血清抗体水平可作为病理标志。例如，与自身免疫性疾病相关的抗体是实验室检测的常规目标之一。简单的临床测试可以提高实验室实践的效果。这项研究描述了一种通过金纳米颗粒的局部表面等离子体共振(LSPR)来光学检测人血清中抗体的一步、免洗技术。作为概念验证实验，基于LSPR的生物传感器使用传统的96孔微滴定板和平板读取器以免洗的方式测量了抗生素在人血清中的溶出量。为了对生物传感器进行有效的表面修饰，将两性离子共聚物作为支架材料固定在金纳米粒子表面，用于固定抗原和封闭剂。成功地实现了人血清中抗生素的一步免洗法测定。此外，由于我们使用的共聚物和亲水性抗原试剂都是由聚环氧乙烷间隔基团组成，因此来自血清内容物的非特异性反应显著减少。血清和缓冲液中的抗原-抗体反应的对比实验表明，血清是生物反应的良好环境。总之，我们基于金纳米颗粒的LSPR方法可以在临床实践中提供一种快速而简单的定量测定血清中抗体含量的方法。图为两种不同的金纳米颗粒表面修饰方法的实验装置：生物素化牛血清白蛋白和两性离子共聚物。用96孔金纳米颗粒固定的微滴定板，通过LSPR光谱的峰位移来测量附着在生物素表面的抗生素的量
In clinical settings, serum antibody levels serve as markers of pathology. For example, antibodies related to autoimmune diseases are among the conventional targets in laboratory tests. Simple clinical tests can improve the efficacy of laboratory practice. This study describes a single-step, wash-free technique for optically detecting antibodies in human serum through the localized surface plasmon resonance (LSPR) of gold nanoparticles. As a proof-of-concept experiment, the amount of antibiotin dissolved in human serum was measured with a LSPR-based biosensor in a wash-free manner using a conventional 96-well microtiter plate and a plate reader. For an efficient surface modification of biosensors, zwitterionic copolymer was used as a scaffold on the gold nanoparticle surface to immobilize antigen and blocking reagent. Single-step, wash-free measurement of antibiotin in human serum was successfully achieved. In addition, nonspecific responses from serum contents were significantly reduced because both the copolymer and hydrophilic antigen reagent that we employed were composed of poly(ethylene oxide) spacer. Comparative experiments of the antigen-antibody reaction in serum to that in buffered solution revealed that serum is a favorable environment for the biological reaction. In conclusion, our gold-nanoparticle-based LSPR method may provide a rapid and simple way to measure the amount of antibody in serum quantitatively in clinical practice.FigureExperimental setups illustrating two different methods of surface modification for the gold nanoparticles: biotinylated BSA and zwitterionic copolymer. The amount of antibiotin that attached to biotin on the surface was measured by the peak shift of LSPR spectra using a 96-well microtiter plate with immobilized gold nanoparticles