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

本计画旨在研制一种可携式之多通道流动池表面电浆共振感测器，利用免疫反应来进行过敏症诊断。特别是，组胺和免疫球蛋白E（IgE）已被证明在人类过敏症的发展中起着非常重要的作用。本研究建立了一种基于表面等离子体共振免疫传感器的组胺和IgE的快速检测方法。此外，我们还开发了一种多通道型表面等离子体共振传感器与小尺寸。本项目取得的研究成果如下。（1）建立了组胺的测定方法。采用硫醇类化合物固定化组胺，再与组胺进行氨基偶联的方法制备了组胺传感器芯片。将含有恒定浓度的抗组胺抗体的样品组胺溶液引入传感器芯片中。抗组胺剂与组胺竞争性反应， ...更多信息 固定在传感器芯片上的共轭物和样品溶液中的组胺。与组胺缀合物结合的所得抗组胺抗体产生SPR传感器的传感器信号。组胺的检测下限为3 ppb。发现传感器芯片可以通过用HCl溶液再生而重复使用。（2）建立了IgE的测定方法。通过物理吸附固定抗IgE（D）抗体，制备IgE传感器芯片。将含有过量抗IgE（H）抗体的样本IgE溶液引入传感器芯片中。将样品溶液中的IgE转化为IgE-抗IgE（H）的免疫复合物，并将所得免疫复合物与固定在传感器芯片上的抗IgE（D）结合。IgE的这种夹心反应产生SPR传感器的大传感器信号。已达到10 ppb IgE的检测下限。(3)A已经开发了具有多通道流动池的便携式表面等离子体共振传感器。传感器的尺寸和重量分别为170（W）X 110（D）X 150（H）mm和约1.5kg。传感器的光学系统由发光二极管（770 nm）、柱面透镜、收集球面透镜、偏振器、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。传感器信号与蔗糖浓度之间存在良好的线性关系，在20 mM的浓度范围内，检测下限为0.3 mM。该值对应于<-6>折射率灵敏度的10 μ RIU水平。少

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