Development of molecular separation / analysis device based on electrokinetic effects

基于动电效应的分子分离/分析装置的开发

• 批准号: 12555076
• 负责人: WASHIZU Masao
• 金额: $ 6.85万
• 依托单位: The University of Tokyo (2002)Kyoto University (2000-2001)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12555076/
• 关键词: Dielectrophoresis; Micro Chemical Analysis System; Micro TAS; Cross-flow injector; Separation; Assay; Dialysis; Micro Fabrication; DNA; クロマトグラフィー; 生体分子; μ-TAS; 電気力学; マイクロマシーニング

The purpose of this investigation is to develop dielectrophoretic chromatography, where high-frequency high- intensity field is created in aqueous solution using microfabricated electrodes, which is utilized for dielectrophoretic separation and assay of biological molecules. An emphasis is laid on the development of prototypes for practical application, so the team consists of researchers from both university and industry. The results obtained can be summarized as below :1) Fabrication process of the electrodes and PDMS-based fluid channels, as well as quantitative on-chip measurement method of molecular concentration based on fluorescence, are developed.2) With the DEP chromatography with a constant-velocity sample flow, B/F (Bound/Free) separation of avidin-biotin binding, DNA hybridization and antigen-antibody binding are experimentally demonstrated. For the DEP chromatography with pulse-wise injection, pressure-driven on-chip cross-injector is used, and the system is shown to be a very fast separation method for DNA larger than several kilo base-pairs. Its applicability to protein separation is also demonstrated.3) A method to integrate dialysis membrane onto the DEP chromatography chip is also developed. The method uses an opening with sharp edge fabricated by anisotropic etching, on which dialyzing polymer membrane is formed by surface tension. The performance of the dialysis membrane is shown to meet the requirement or the sample pre-treatment for the DEP chromatography.

本研究的目的是开发介电泳色谱，其中使用微加工电极在水溶液中产生高频高强度场，用于介电泳分离和生物分子的分析。重点放在实际应用的原型开发上，因此该团队由来自大学和工业的研究人员组成。主要研究结果如下：1）建立了基于PDMS的流体通道和电极的制备工艺，以及基于荧光的分子浓度定量测量方法; 2）采用恒速样品流的DEP色谱，实现了亲和素-生物素结合、DNA杂交和抗原-抗体结合的B/F（Bound/Free）分离。对于脉冲进样的DEP色谱，使用压力驱动的芯片上交叉进样器，该系统被证明是一种非常快速的DNA分离方法大于几千个碱基对。3）提出了一种将透析膜集成到DEP色谱芯片上的方法。该方法利用各向异性腐蚀制作一个具有尖锐边缘的开口，在开口上通过表面张力形成透析聚合物膜。结果表明，该透析膜的性能满足DEP色谱对样品前处理的要求。

项目成果

Tomohisa Kawabata and Masao Washizu: "Dielectrophoretic Detection of Molecular Bindings"IEEE Transaction IA. 37, No.6. 1625-1633 (2001)

Tomohisa Kawabata 和 Masao Washizu：“分子结合的介电泳检测”IEEE Transaction IA。

Hiroyuki Kabata,Wataru Okada,Masao Washizu: "Single-Molecule Dynamics of the Eco RI Enzyme using Stretched DNA : Its Application to In Situ Sliding Assay and Optical DNA Mapping"Jpn.J.Appl.Phys.. Vol.39. 7164-7171 (2000)

Hiroyuki Kabata、Wataru Okada、Masao Washizu：“使用拉伸 DNA 的 Eco RI 酶的单分子动力学：其在原位滑动测定和光学 DNA 绘图中的应用”Jpn.J.Appl.Phys. 第 39 卷。

軍司昌秀, T.B.Jones, 鷲津正夫: "誘電泳動による液滴の形成と融合"静電気学会2002全国大会講演論文集. 155-158 (2002)

Masahide Gunji、T.B.Jones、Masao Washizu：“介电泳液滴的形成和融合”静电学会 2002 年全国会议记录 155-158 (2002)。

M.Gunji, T.B.Jones, M.Washizu: "DEP-driven simultaneous 2 x 2 droplet mixer array"Proc. Micro total analysis systems 2002. 721-723 (2002)

M.Gunji、T.B.Jones、M.Washizu：“DEP 驱动的同时 2 x 2 液滴混合器阵列”Proc。

Masao Washizu: "Bio-Nanotechnology of DNA"IEEE-EMBS Special Topic Conference on Microtechnologies in Medicine and Biology Proceedings. 1-6 (2002)

Masao Washizu：“DNA 生物纳米技术”IEEE-EMBS 医学和生物学微技术专题会议。