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利用光学微芯片生物传感器进行无标记的抗体-抗原相互作用监测。

Label-free monitoring of antibody-antigen interactions using optical microchip biosensors.

机构信息

Stratophase Ltd., Unit 10a, The Quadrangle, Premier Way, Romsey SO51 9DL, United Kingdom.

出版信息

J Immunol Methods. 2010 Oct 31;362(1-2):121-6. doi: 10.1016/j.jim.2010.09.015. Epub 2010 Sep 17.

DOI:10.1016/j.jim.2010.09.015
PMID:20850447
Abstract

A rapid, label-free optical biosensor system for sensitive monitoring of bio-molecular interactions in real-time is presented. SpectroSens sensor chips are based on integrated planar Bragg gratings sensitive to localised changes in refractive index. Bio-molecule recognition is imparted by functionalisation of the sensing surface with antibodies against targets of interest. In this study, antibodies against selected proteins were oriented with recombinant Protein A/G, which was covalently immobilised to the sensor chip via amine coupling to a glutaraldehyde-activated silane layer. Immunoassays for the detection of rabbit IgG and ovalbumin proteins as model antibody-antigen interaction systems were performed. Binding of complementary antigens to respective antibody-functionalised sensors manifested as changes in wavelength of light reflected from the optical sensors. Quantitative binding kinetics with detection sensitivities in the mid ng/ml range were obtained for both antigens using this planar, two-dimensional surface coating. Data presented demonstrate the suitability of SpectroSens sensors as a valuable tool in life science research and development for monitoring bio-specific interactions, protein concentration determination and antibody selection; the optical integration and analytical characteristics of these sensors suggest that they may find numerous applications in bio-pharmaceutical development and clinical diagnostics.

摘要

一种快速、无标记的光学生物传感器系统,用于实时敏感监测生物分子相互作用,被提出来了。SpectroSens 传感器芯片基于对局部折射率变化敏感的集成平面布拉格光栅。通过用针对感兴趣的靶标的抗体对传感表面进行功能化,赋予生物分子识别功能。在这项研究中,针对选定蛋白质的抗体通过重组蛋白 A/G 定向,该蛋白通过与戊二醛激活的硅烷层的胺偶联而共价固定在传感器芯片上。进行了兔 IgG 和卵清蛋白蛋白作为模型抗体-抗原相互作用系统的免疫测定。互补抗原与各自抗体功能化传感器的结合表现为从光学传感器反射的光的波长变化。使用这种平面二维表面涂层,两种抗原都获得了 mid ng/ml 范围内的定量结合动力学和检测灵敏度。所呈现的数据表明 SpectroSens 传感器适合用作生命科学研究和开发中的有价值工具,用于监测生物特异性相互作用、蛋白质浓度测定和抗体选择;这些传感器的光学集成和分析特性表明,它们可能在生物制药开发和临床诊断中找到许多应用。

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