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

本项目的目的是研制一种便携式多通道流通池表面等离子体共振传感器，利用免疫反应进行变态反应诊断。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过程中起着非常重要的作用。在这个项目中，我们开发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，我们还研制了一种体积较小的多道型表面等离子体共振传感器。本课题取得的研究成果如下：(1)建立了组胺的测定方法。将烷基硫醇类化合物固定化，再与组胺进行氨基偶联，制备了组胺传感器芯片。将含有恒定浓度的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…的竞争性反应传感器芯片和样品溶液中组胺的固定化程度较高。得到的抗组胺抗体结合到组胺结合物上，产生SPR传感器的传感器信号。该方法对组胺的检出限为3 ppb。该传感器芯片经盐酸溶液再生后可重复使用。(2)建立了检测IgE的方法。用物理吸附的方法固定化抗IgE(D)抗体，制备了一种检测IgE的传感器芯片。将含有过量抗IgE(H)抗体的样品IgE溶液引入传感器芯片。样品溶液中的免疫球蛋白被转化为IgE-抗-IgE(H)的免疫复合物，得到的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了SPR传感器的大量传感器信号。检测下限可达10 ppb。(3)研制了一种便携式多通道流通池表面等离子体共振传感器。传感器的尺寸和重量分别为170(宽)×110(深)×150(高)毫米和约1.5公斤。该传感器的光学系统由发光二极管(770 Nm)、柱面透镜、集合球透镜、偏振器、反射镜和线阵CCD传感器组成。利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在20 mM范围内呈良好的线性关系。检测下限为0.3 mm。该值对应于折射率灵敏度的10^lt；-6 RIU水平。较少

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