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Research on micro environmental analysis using stacked miniaturized 3D chemical system

堆叠式微型3D化学系统微环境分析研究

基本信息

批准号：
16550083
负责人：
UTSUMI Yuichi
金额：
$ 2.3万
依托单位：
University of Hyogo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16550083/
关键词：
environmental analysis on chip analysis bio analysis micro mechanics bio reactor reaction flow

项目摘要

We proposed and fabricated a new chemical reactor with a vertical fluid flow operation for the wide biochemical applications. For rapid and high yield reaction between reagents with large quantity, the filters, which possess 2,100-cylindrical through-bores with 40pm diameter in 200pm-thickness PMMA sheets, were fabricated by deep X-ray lithography using synchrotron-radiation from LIGA beamline BL2 of NewSUBARU. We tried the application of the fabricated vertical reactor to an sandwich enzyme-linked immunosorbent assay (ELISA) for the analysis of mouse immunoglobulin (IgG) and nonylphenol using ultra violet (UV) absorption spectroscopy. We succeeded to obtain calibration curve of Mouse IgG and found that the mouse IgG less than 100 ng/ml concentration was quantitatively detected within 45 min of total analytical period, which was about one third of the period required for the conventional method using micro titer plate. We investigated the possibility of the analysis methods using the proposed vertical micro reactor for competitive enzyme-linked immunosorbent assay (ELISA), which enable high sensitive detection of an endocrine disrupter nonylphenol in a series of vertical fluidic operation. In the assay using filters pretreated with 0.3 Dg/ml of anti-NP-antibody, even at 0.01ng/m1 of free NP, we still observed the inhibition of the binding (B/B_0=90%). Under this condition, calibration curve of NP was obtained at the range of 0.01-10ng/ml. This sensitivity was two orders higher than the sensitivity (5-100 ng/ml) obtained by ordinary ELISA using 96-wells-micro-titer plate and the same anti-NP-antibody.

我们提出并制作了一种新型的垂直流化学反应器，用于广泛的生物化学应用。为了在大量试剂之间进行快速和高产率的反应，使用NewSUBARU的LIGA光束线BL 2的同步辐射通过深X射线光刻制造了在200 μ m厚的PMMA板中具有2，100个直径为40 μ m的圆柱形通孔的过滤器。我们尝试了应用所制造的垂直反应器的夹心酶联免疫吸附测定（ELISA）的小鼠免疫球蛋白（IgG）和壬基酚的分析，使用紫外（UV）吸收光谱。成功地获得了小鼠IgG的校准曲线，发现小于100 ng/ml浓度的小鼠IgG在45 min的总分析周期内被定量检测到，约为使用微量滴定板的常规方法所需时间的三分之一。我们调查的分析方法的可能性，使用拟议的垂直微反应器的竞争性酶联免疫吸附测定（ELISA），这使得高灵敏度检测的内分泌干扰物壬基酚在一系列的垂直流体操作。在使用用0.3 μ g/ml抗NP抗体预处理的滤膜的测定中，即使在0.01ng/ml游离NP下，我们仍然观察到结合的抑制（B/B_0=90%）。在此条件下，NP的线性范围为0.01 ~ 10 ng/ml。此灵敏度比用96孔微量滴定板和相同的抗NP抗体的普通ELISA获得的灵敏度（5-100 ng/ml）高两个数量级。

项目成果

期刊论文数量（44）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Vertical Micro Reactor Stack for Integrated Chemical Reaction System

用于集成化学反应系统的立式微反应堆

DOI：
发表时间：
2005
期刊：
e-Journal of Surface Science and Nanotechnology 13
影响因子：
0
作者：
T.Asano;Y.Ukita;K.Matsui;M.Takeo;S.Negoro;Y.Utsumi
通讯作者：
Y.Utsumi

Fabrication of a Micro Reactor for Vertiocal Unit Operation using Multifunctional Fluid Filter and its Application to Biochemical Reaction

多功能流体过滤器垂直单元操作微反应器的制作及其在生化反应中的应用

DOI：
发表时间：
2007
期刊：
Microsystem Technologies, 13
影响因子：
0
作者：
T Asano;Y.Ukita;K.Matsui;M.Takeo;S.Negoro;Y.Utsumi
通讯作者：
Y.Utsumi

Enzyme-Linked Immunosorbent Assay Using Vertical Micro Reactor Stack for the Detection of Bio molecules

使用立式微反应堆进行酶联免疫吸附测定来检测生物分子

DOI：
发表时间：
2007
期刊：
IEEJ Transactions on Electronics, Information and Systems (in Japanese) 247, 2
影响因子：
0
作者：
K.Matsui;T.Ohmura;S.Morimoto;T.Asano;Y.Ukita;Y.Utsumi;M.Takeo;S.Negoro
通讯作者：
S.Negoro

Novel Characteristics of Multifunctional Fluid Filters Fabricated by High-Energy Synchrotron Radiation Lithography

高能同步辐射光刻制备多功能流体过滤器的新特性