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个新的化学反应器，并为广泛的生化应用提供了垂直流体流动的操作。对于大量量的试剂之间的快速和高产量反应，使用Deep X射线光刻使用Synchrotron-radiation从Newsubaru的Liga Beamline bl2制造了深层X射线光刻，该过滤器在200pm厚度PMMA板上具有2100列透透透过孔，直径为40pm，直径为40pm。我们尝试使用Ultra Violet（UV）吸收光谱法应用了制造的垂直反应器在三明治酶联免疫吸附测定（ELISA）中用于分析小鼠免疫球蛋白（IgG）和Nonylphenol的分析。我们成功获得了小鼠IgG的校准曲线，发现小鼠IgG在总分析周期的45分钟内定量检测到小于100 ng/ml的浓度，这约为使用微滴度板所需的传统方法所需的三分之一。我们研究了使用拟议的垂直微反应器进行分析方法的可能性，以进行竞争性酶联免疫吸附测定法（ELISA），该测定能够在一系列垂直流体运行中对内分泌偏离甲醇的高敏感检测。在使用0.3 dg/mL抗NP抗体预处理的过滤器的测定中，即使在自由NP的0.01ng/m1下，我们仍然观察到结合的抑制作用（B/B_0 = 90％）。在这种情况下，在0.01-10ng/ml的范围内获得了NP的校准曲线。该灵敏度比普通ELISA使用96孔微滴度板和相同的抗NP抗体获得的敏感性（5-100 ng/ml）高两个阶。

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