| Project/Area Number |
17310087
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | Nagoya University (2006-2007)
Kanagawa Academy of Science and Technology (2005) |
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Principal Investigator |
MANABU Tokeshi Nagoya University, Graduate School of Engineering, Associate Professor (60311437)
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| Co-Investigator(Kenkyū-buntansha) |
KITAMORI Takehiko The University of Tokyo, Graduate School of Engineering, Professor (60214821)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥15,230,000 (Direct Cost: ¥14,300,000、Indirect Cost: ¥930,000)
Fiscal Year 2007: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2006: ¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 2005: ¥6,300,000 (Direct Cost: ¥6,300,000)
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| Keywords | Microchip / Immunoassay / Photopolymer / Fluorescence Polarization / マイクロ化学チップ / 酵素免疫分析 / 抗原抗体反応 / Protein A |
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| Research Abstract |
We have developed three types of new immunoassay formats.
A microchip-based semi-automated heterogeneous immunoassay was developed wherein a flow injection-like system and repeatable-use gel were employed. A regeneration buffer for antigen-antibody interaction of brain natriuretic peptide (BNP) was found which did not lose binding activity of the antibody on the gels and integration of such an immunoassay scheme into a semi-automated microfluidics system would facilitate repeated-use heterogeneous immunoassay. The concentration of BNP in the range of 0.1-100 pg/ml was successfully determined using the system and protocol developed here.
We developed a new platform for chip-based immunoassay to create the effective reaction sites of immunological reaction between antigen and antibody. The reaction sites were fabricated by the UV photopolymerization of a solution of photopolymer and the antibody-immobilized polystyrene beads. Our platform has many advantages ; (i) rapid analysis (a few min
… More
utes), (ii) easy handling (no need pump and valve), (iii) low-cost (disposable), (iv) small volume(I droplets(0.25gL)of sample and reagents), and(v)high sensitivity(2 orders higher than conventional method). In order to evaluate our platform, we carried out(sandwich)immunoassay for a-fetoprotein(AFP), which is known as a tumor marker. In the case of 1-min incubation(total assay time : ca. 3 min), the limit of detection (LOD) was 10 ng/ml. This value is compared to that of conventional method using a microtiter plate with 4 h of total assay time. The LOD for &min incubation was 2 orders lower than that of conventional one.
We developed an optimized microchip-based fluorescence polarization immunoassay (FPIA) . Microchip-based FPIA was simple and rapid because it did not require several processes such as washing and reflowing and immobilization of antibodies or antigens in a channel. The quantitative analysis of theophylline was performed in short analysis time (65 seconds) with simple operation and reduced samples. The microchip-based FPIA will find a frequent use as point-of-care testing in clinical field. Furthermore we observed the behavior of fluorescence polarization after antigens and antibodies are mixed in a microchannel. The information will lead to the development of immunoassays using fluorescence polarization. Less
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