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

• 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

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.

我们提出并制造了一种具有垂直流体流动操作的新型化学反应器，用于广泛的生化应用。为了实现大量试剂之间的快速、高产率反应，过滤器采用深度 X 射线光刻技术，使用来自 LIGA 光束线 BL2 的同步辐射，在 200μm 厚度的 PMMA 片上具有 2,100 个直径为 40μm 的圆柱形通孔。新斯巴鲁。我们尝试将组装的立式反应器应用于夹心酶联免疫吸附测定 (ELISA)，使用紫外线 (UV) 吸收光谱分析小鼠免疫球蛋白 (IgG) 和壬基酚。我们成功获得了小鼠 IgG 的校准曲线，发现在总分析时间 45 分钟内即可定量检测浓度低于 100 ng/ml 的小鼠 IgG，大约是使用微量滴定板的传统方法所需时间的三分之一。我们研究了使用所提出的立式微反应器进行竞争性酶联免疫吸附测定（ELISA）的分析方法的可能性，该分析方法能够在一系列立式流体操作中高灵敏度地检测内分泌干扰物壬基酚。在使用用 0.3 Dg/ml 抗 NP 抗体预处理的过滤器进行的测定中，即使在 0.01ng/m1 的游离 NP 下，我们仍然观察到结合的抑制 (B/B_0=90%)。在此条件下，得到0.01-10ng/ml范围内NP的校准曲线。该灵敏度比使用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（5-100ng/ml）高两个数量级。

项目成果

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

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

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

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

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

• 发表时间: 2006
• 期刊: Japanese Journal of Applied Physics 45, 9A
• 作者: Y.Ukita;T.Asano;Y.Utsumi
• 通讯作者: Y.Utsumi

A New Micro-Chemical Reactor Using Vertical Fluid Flow Operation

一种采用垂直流体流动操作的新型微化学反应器

• 发表时间: 2005
• 期刊: Japanese Journal of Applied Physics 44
• 作者: Y.Utsumi;T.Asano;Y.Ukita;K.Matsui;M.Takeo;S.Negoro
• 通讯作者: S.Negoro

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

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