| Project/Area Number |
16510100
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Microdevices/Nanodevices
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| Research Institution | National Institute of Advanced Industrial Science and Technology |
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Principal Investigator |
SHIMOSAKA Takuya National Institute of Advanced Industrial Science and Technology, National Metrology Institute of Japan, Researcher, 計測標準研究部門, 研究員 (40295473)
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| Co-Investigator(Kenkyū-buntansha) |
UCHIYAMA Katsumi Tokyo Metropolitan University, School of Engineering, Associate Professor, 工学研究科, 准教授 (40151899)
NAKAGAWA Tatsuro Tokyo Metropolitan University, School of Engineering, Associate Professor, 工学研究科, 准教授 (50244421)
KATO Kenji National Institute of Advanced Industrial Science and Technology, National Metrology Institute of Japan, Senior Researcher, 計測標準研究部門, 科長 (00356784)
前田 恒昭 東京本部, ベンチャー開発戦略研究センター, 室長 (00357980)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2005: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2004: ¥2,700,000 (Direct Cost: ¥2,700,000)
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| Keywords | micro-TAS / ELISA / enzyme / antibody / PDMS / photo-cross linking / regioselective modification / マイクロチップ / 抗原抗体反応 / 酵素反応 |
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| Research Abstract |
The micro channel made of PDMS was used as an enzyme and/or antigen-antibody (Ag/Ab) reaction chamber. The inner surface of the channel was region-selectively immobilized with enzyme and/or antibody. We applied the microchip to enzymatic glucose sensor and enzyme immunoassay. One of the applications was ELISA on the microchip. We developed an ELISA system on the micro channel, and then the method was compared with a conventional ELISA measurement, that is, ELISA using a micro titer plate with 96-holes array. First antibody was immobilized on the micro channel and then the channel was blocked with BSA and/or casein for the prevention from non-specific adsorption. Then analyte IgA and second antibody which was previously labeled with Horse Radish Peroxidase (HRP), was added to the micro channel. Finally Amplex Red (AR) solution was added to the channel. Concentration of IgA was determined through the measurement of fluorescence of Resorufin. Calibration curve for IgA by the method showed good linearity in the range 0〜250 ng/mL with the following conditions ; Ag/Ab reaction time ; 1.4 min, enzyme reaction with AR ; 0.1min. We did same kinds of experiments by the conventional ELISA method, and measuring time of the micro chip method was about 60-times faster than that of the conventional method. The concentration of real human saliva sample obtained by both methods agreed well.
The second application was enzymatic glucose sensor, in which region-selective enzyme modification technique was used. Glucose oxidase and HRP were region-selectively immobilized on separate position of the inner surface of the micro channel. Glucose concentration was determined by the channel. Linear relationship between the concentration of glucose and the fluorescence intensity was obtained in the range 8〜130 μM. Reaction time was about five seconds, which was about 370 times faster than the conventional method with micro titer plate.
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