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

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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