Reaction, Separation and Dtection of Biomacromolecules using Disposable Microchips

• Project/Area Number: 11450318
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 生物・生体工学
• Research Institution: The University of Tokyo
• Principal Investigator: FUJII Teruo The University of Tokyo, Institute of Industrial Science, Associate Professor (30251474)
• Co-Investigator(Kenkyū-buntansha): SEKI Minoru 東京大学, Dept of Chemistry and Biotechnolgy Graduate school of Engineering, Assosiate Professor (80206622) ; YAMAMOTO Takatoki 東京大学, Institute of Industrial Science, Assistant (20322688)
• Project Period (FY): 1999 – 2001
• Project Status: Completed (Fiscal Year 2001)
• Budget Amount: ¥5,500,000 (Direct Cost: ¥5,500,000); Fiscal Year 2001: ¥1,900,000 (Direct Cost: ¥1,900,000); Fiscal Year 2000: ¥3,600,000 (Direct Cost: ¥3,600,000)
• Keywords: Microchip / Silicon / Electrophoreisis / PCR (Polymerase chain reaction) / Cell-free Protein Synthesis / キャピラリ電気泳動 / ディスポーザブルチップ / Micro-TAS / DNA解析 / ゲノム解析 / キャピラリー電気泳動 / μ-TAS

Research Abstract

In this research project, we have been trying to realize low-cost microchips for biochemical reaction and analysis, which can be fabricated through micro-molding process of the silicone rubber material using silicon wafer as a mold master. Based on the research progress achieved in the previous years, we investigated the performance of the microchip for free-flow Electrophoretic Separation of DNA fragments in detail. And on-chip biochemical reactions such as PCR (polymerase chain reaction) and cell-free protein synthesis were also conducted to examine the potentials of the developed microchip.
1) Free-flow Electrophoresis: Cr-Au electrodes are patterned on a glass substrate for application of high voltage electric field required for the electrophoretic separation of DNA fragment. By pasting the silicone rubber microchip, which has microfluidic structure on it, on the glass substrate, we could carry out the experiments on electrophoresis of DNA fragments. 100bp to several hundreds by DNA fragments can be successfully separated and detected within just two or three minutes. This is almost ten to fifteen times faster than the conventional slab-gel based method.
2) Biochemical Reactions: with the same hybrid structure with a glass substrate and a silicone rubber microchip, we have conducted two kinds of biochemical reactions with temperature control through the integrated electrodes; PCR and cell-free protein synthesis. For the PCR reaction , it is confirmed that 700bp DNA fragment can be amplified. For cell-free protein synthesis, it is proved that Green Fluorescent Protein can be synthesized by detecting the emitted fluorescence from the target molecule.

Research Products

• [Journal Article] Microfabricated Polymer Chip for Capillary Gel Electrophoresis (2001)
  • Author(s): Hong,J.-W., Hosokawa, K., Fujii, T., Seki, M., Endo, I.
  • Journal Title: Biotechnology Progress Vol. 17, Issue 5 Pages: 958-962
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Development of PDMS-based Microbiochemical Systems (2001)
  • Author(s): Fujii, T
  • Journal Title: Program & Abstracts, IUPAC International Conference on Analytical Sciences 2001, Tokyo 3A12 Pages: 80-80
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] PDMS-based Microfluidic Devices for Biomedical Applications (2001)
  • Author(s): Fujii, T
  • Journal Title: Abstracts of Micro & Narro Engineering 2001 Conference, Grenoble Pages: 78-78
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cell-free Protein Synthesis in PDMS-based Parallel Microreactors (2001)
  • Author(s): Yamamoto, T., Nojima, T., Fujii, T
  • Journal Title: Proceedings of Micro-TAS2001, Monterey CA Pages: 69-77
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Separation and Collection of a Specified DNA Fragment by Chip-based CE System (2001)
  • Author(s): Hong, J.-W., Hagiwara, H., Fujii, T., Machida, H., Inoue, M., Seki, M., Endo, I
  • Journal Title: Proceedings of Micro-TAS2001, Monterey CA Pages: 113-114
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Voltage Control for Separation and Collection of a Specified Fragment on a Chip (2001)
  • Author(s): Hong, J.-W., Hagiwara, H., Fujii, T, Machida, H., Inoue, M., Seki, M., Endo, I
  • Journal Title: 第21回キャビラリー電気泳勤シンポジウム要旨集、神戸
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Microfabricated Polymer Chip for Capillary Gel Electrophoresis (2001)
  • Author(s): Hong, J.-W., Hosokawa, K., Fujii, T., Seki, M., Endo, I.
  • Journal Title: Biotechnology Progress Vol.17, Issue 5 Pages: 958-962
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Development of PDMS-based Microbiochemical Systems (2001)
  • Author(s): Fujii, T.
  • Journal Title: Program & Abstracts, 1UPAC International Conference on Analytical Sciences 2001, Tokyo 3A12 Pages: 80-80
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] PDMS-based Microfluidic Devices for Biomedical Applications (2001)
  • Author(s): Fujii, T.
  • Journal Title: Abstracts of Micro & Nano Engineering 2001 Conference, Grenoble Pages: 78-78
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cell-free Protein Synthesis in PCOS-based Parallel Microreactors (2001)
  • Author(s): Yamamoto, T., Nojima, T., Fujii, T.
  • Journal Title: Proceedings of the 5th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (Micro-TAS2001) Monterey CA Pages: 69-71
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Separation and Collection of a Specified DNA Fragment by Chip-based CE System (2001)
  • Author(s): Hong, J.-W., Hagiwara, H., Fujii, T., Machida, H., Inoue, M., Seki, M., Endo, I.
  • Journal Title: Proceedings of the 5th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (Micro-TAS 2001) Monterey CA Pages: 113-114
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Voltage Control for Separation and Collection of a Specified Fragment on a Chip (2001)
  • Author(s): Hong, J.-W., Hagiwara, H., Fujii, T., Machida, H., Inoue, M., Seki, M., Endo, I.
  • Journal Title: 第21回キャピラリー電気泳動シンポジウム要旨集、神戸
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Hong, J.-W., Hosokawa, K., Fujii, T., Seki, M., Endo, I.: "Microfabricated Polymer Chip for Capillary Gel Electrophoresis"Biotechnology Progress. Vol.17, Issue 5. pp.958-pp.962 (2001)
• [Publications] Fujii, T.: "Development of PDMS-based Microbiochemical Systems"Program & Abstracts, IUPAC International Conference on Analytical Sciences 2001, Tokyo. 3A12. 80 (2001)
• [Publications] Fujii, T.: "PDMS- based Microfluidic Devices for Biomedical Applications"Abstracts of Micro & Nano Engineering 2001 Conference, Grenoble. 78 (2001)
• [Publications] Yamamoto, T., Nojima, T., Fujii, T.: "Cell-free Protein Synthesis in PDMS-based Parallel Microreactors"Proceedings of Micro-TAS 2001, Monterey CA. pp.69-pp.71 (2001)
• [Publications] Hong, J.-W., Hagiwara, H., Fujii, T., Machida, H, , Inoue, M., Seki, M., Endo, I.: "Separation and Collection of a Specified DNA Fragment by Chip-based CE System"Proceedings of Micro-TAS 2001, Monterey CA. pp.113-pp.114 (2001)
• [Publications] Hong, J.-W., Hagiwara, H., Fujii, T., Machida, H., Inoue, M., Seki, M., Endo, I.: "Voltage Control for Separation and Collection of a Specified Fragment on a Chip"第21回キャピラリー電気泳動シンポジウム要旨集、神戸.
• [Publications] Hong,J.W.,Fujii,T.,Seki,M.,and Endo,I.: "Optimization of DNA Separation by Capillary Gel Electrophoresis on a Microfabricated Polymer Chip"Chemical Engineering Symposium Series. Vol.65. 177-180 (2000)
• [Publications] Hong,J.W.,Hosokawa,K.,Fujii,T.,Seki,M.,and Endo,I.: "Microfabricated Structures for Bioseparation"Bioseparation Engineering, Progress in Biotechnology, Elsevier. 16. 69-74 (2000)
• [Publications] 藤井輝夫,福場辰洋,長沼毅,遠藤勲: "マイクロチップによる生化学反応分析技術の開発と極限環境微生物探査への応用"第1回極限環境微生物学会年会講演要旨集. 27-30 (2000)
• [Publications] Fujii,T.: "Downsizing Biochemical/Chemical Processes-Technologies and Future Perspectives-"Proceedings of the International Workshop on Microfactories (IWMF2000). 111-114 (2000)
• [Publications] Fujii,T.: "New Approach to Chemistry and Biochemistry through Microchip Technology"Proceedings of the Sixth International Micromachine Symposium. 105-109 (2000)
• [Publications] Hong,J.W.,藤井輝夫,関実,山本貴富喜,遠藤勲: "PDMS.ガラスハイブリッドマイクロチップによる遺伝子解析"第20回キャピラリー電気泳動シンポジウム要旨集. 48-49 (2000)
• [Publications] Houg,J.W.,他: "An Inexpensive PDMS Microchip for Capillary Electrophoresis"Proc.Transducers'99. 760-763 (1999)
• [Publications] Houg,J.W.,他: "Bioseparation of Biomolecules in Microfabricated Structures"Proc.RRR'99. (1999)
• [Publications] Seki,M.,他: "μBioChemical Engineering"Proc.YABEC'99. 13-15 (1999)
• [Publications] 藤井輝夫: "マイクロ生化学システムの開発"第4回バイオ・ナノエレクトロニクス研究センターシンポジウム講演集. 8-17 (1999)
• [Publications] 藤井輝夫: "ディスポーザブル電気泳動チップとその周辺技術"第19回キャピラリー電気泳動シンポジウム講演要旨集. 18-19 (1999)
• [Publications] Hosokawa K.,Fujii T.and Endo I.: "Handling of Picoliter Liquid Samples in a PDMS-based Microfluidic Device"Analytical Chemistry. 71・20. 4781-4785 (1999)
• [Publications] 藤井輝夫、他: "マイクロリアクター -技術の現状と展望-"住化技術情報センター. 241 (1999)