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Development of multi-channel type surface plasmon resonance sensor and its application to simultaneous determination of allergy related substances

多通道型表面等离子体共振传感器的研制及其在过敏相关物质同时测定中的应用

基本信息

批准号：
16350047
负责人：
IMATO Toshihiko
金额：
$ 10.11万
依托单位：
KYUSHU UNIVERCITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16350047/
关键词：
surface plasmon resonance allergy immunoassay microchannel competitive assay サンドイッチ測定法

项目摘要

The propose of this project is to develop a portable surface plasmon resonance sensor with a multi-channel flow cell for allergy diagnosis by using immunoreactions. Especially, histamine and immunoglobulin E (IgE) have proven to play a very important role in the development of allergies in humans. In this project, we have developed a simple and rapid method for the determination of histamine and IgE based on a surface plasmon resonance immunosensor. In addition, we have developed a multi-channel type surface plasmon resonance sensor with a small size. The research results obtained in this project is as follows.(1)The determination method for histamine has been achieved. A sensor chip for histamine was prepared by immobilization of alkane thiol compounds followed by amino-coupling with histamine. A sample histamine solution containing an anti-histamine antibody at a constant concentration was introduced into the sensor chip. The anti-histamine reacts competitively with the histamine con … More jugate immobilized on the sensor chip and histamine in the sample solution. The resulting anti-histamine antibody bound to the histamine conjugate generated the sensor signal of the SPR sensor. A lower detection limit of 3 ppb histamine was achieved by the proposed method. The sensor chip was found to be possible to use repeatedly by regeneration with an HC1 solution.(2)The determination method for IgE has been achieved. A sensor chip for IgE was prepared by immobilization of an anti-IgE(D) antibody by physical adsorption. A sample IgE solution containing an excess amount of an anti-IgE(H) antibody was introduce into a sensor chip. IgE in the sample solution was converted to an immunocomplex of IgE-anti-IgE(H) and the resulting immunocomplex is bound to the anti-IgE(D) immobilized on the sensor chip. This sandwich reaction of IgE generated a large sensor signal of the SPR sensor. A lower detection limit 10 ppb IgE has been achieved.(3)A portable surface plasmon resonance sensor with a multi-channel flow cell has been developed. The size and weight of the sensor was 170(W) x 110(D) x 150(H) mm and about 1.5 kg, respectively. The optical system of the sensor consisted of a light emitting diode (770 nm), cylindrical lens, collection spherical lens, polarizer, reflection mirror and a linear CCD sensor. The performance of the sensor was evaluated by using sucrose solution. A good linear relationship between the sensor signal and the concentration of sucrose was found in the concentration range up to 20 mM. The lower detection limit of 0.3 mM was achieved. This value corresponds 10^<-6> RIU level in refractive index sensitivity. Less

本课题拟研制一种便携式多通道流细胞表面等离子体共振传感器，用于免疫反应诊断过敏。特别是，组胺和免疫球蛋白E （IgE）已被证明在人类过敏的发展中起着非常重要的作用。在本项目中，我们建立了一种基于表面等离子体共振免疫传感器的简单快速测定组胺和IgE的方法。此外，我们还开发了一种小尺寸的多通道型表面等离子体共振传感器。本课题的研究成果如下：(1)建立了组胺的测定方法。以烷烃硫醇类化合物为固定载体，氨基偶联组胺，制备了组胺传感器芯片。将含有恒定浓度抗组胺抗体的组胺溶液样品引入传感器芯片。抗组胺药与组胺药竞争性反应，更多的是固定在传感器芯片上和样品溶液中的组胺药。由此产生的抗组胺抗体与组胺偶联物结合，产生SPR传感器的传感器信号。该方法组胺的检出限较低，为3 ppb。发现传感器芯片可以通过HC1溶液再生来重复使用。(2)建立了IgE的测定方法。采用物理吸附法固定抗IgE(D)抗体，制备了IgE传感器芯片。将含有过量抗IgE(H)抗体的IgE溶液样品引入传感器芯片。将样品溶液中的IgE转化为IgE-anti-IgE(H)的免疫复合物，该免疫复合物与固定在传感器芯片上的anti-IgE(D)结合。IgE的这种夹心反应产生了SPR传感器的大传感器信号。较低的检出限已达到10 ppb IgE。(3)研制了一种便携式多通道流池表面等离子体共振传感器。传感器的尺寸为170(W) × 110(D) × 150(H) mm，重量约为1.5 kg。传感器的光学系统由发光二极管（770nm）、圆柱透镜、集合球透镜、偏光片、反射镜和线阵CCD传感器组成。用蔗糖溶液对传感器的性能进行了评价。在20 mM以内的浓度范围内，传感器信号与蔗糖浓度呈良好的线性关系，检测下限为0.3 mM。该值对应于折射率灵敏度的10^<-6> RIU水平。少