Single molecule detection of protein/DNA aptamer complex using giant biological nanpores

• Project/Area Number: 25708024
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Analytical chemistry
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: KAWANO Ryuji 東京農工大学, 工学(系)研究科(研究院), 准教授 (90401702)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥25,740,000 (Direct Cost: ¥19,800,000、Indirect Cost: ¥5,940,000); Fiscal Year 2014: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000); Fiscal Year 2013: ¥16,510,000 (Direct Cost: ¥12,700,000、Indirect Cost: ¥3,810,000)
• Keywords: 電気化学 / 一分子計測 / チャネル膜タンパク質 / 脂質二分子膜 / マイクロ加工 / ナノポア / アプタマー

Outline of Final Research Achievements

We have developed stable lipid bilayer using droplet contact method. This method allows us to measure current signals of larger-sized nanopore proteins with the portable system. Using this system, we are able to measure a serine protease and a toxin protein with DNA aptamer using the large nanopore at the single molecule level.

Research Products

• [Journal Article] A Portable Lipid Bilayer System for Environmental Sensing with a Transmembrane Protein (2014)
  • Author(s): Ryuji Kawano, Yutaro Tsuji, Koki Kamiya, Taiga Kodama, Toshihisa Osaki, Norihisa Miki, and Shoji Takeuchi
  • Journal Title: PLOS ONE Volume: 9 Issue: 7 Pages: e102427-e102427
  • DOI: 10.1371/journal.pone.0102427
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Extend The Size of Biological Nanopore Using Mgainin and Perforin Pores (2014)
  • Author(s): H. Watanabe and R. Kawano
  • Journal Title: Proceedings of MicroTAS Volume: 1 Pages: 2125-2127
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] MICRO-SCALE DROPLET CONTACT METHOD BY MECHANICAL MOTION: REPRODUCIBLE AND ROBUST LIPID BILAYER FORMATION (2013)
  • Author(s): L.N.S. Zaleha, R. Kawano, H. Yasuga, K. Kamiya,T. Osaki, N. Miki and S. Takeuchi
  • Journal Title: Proceedings of Micro TAS 2013 Volume: 1 Pages: 1433-1435
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Stable Lipid Bilayer Formation Using Microfabrication and Its Applications (2015)
  • Author(s): R. Kawano
  • Organizer: 2nd CMS International Symposium
  • Place of Presentation: 九州大学（福岡）
  • Year and Date: 2015-01-20
  • Invited
• [Presentation] Extend The Size of Biological Nanopore Using Mgainin and Perforin Pores (2014)
  • Author(s): H. Watanabe and R. Kawano
  • Organizer: The Seventeenth International Conference on Miniaturized Systems for Chemistry and Life Sciences
  • Place of Presentation: Texas, US
  • Year and Date: 2014-10-26 – 2014-10-30
• [Presentation] マガイニンとパーフォリンを用いたナノポアのサイズの拡張とナノポアセンシングへの応用 (2014)
  • Author(s): 渡辺寛和、川野竜司
  • Organizer: 第4 回CSJ 化学フェスタ
  • Place of Presentation: タワーホール船橋（千葉）
  • Year and Date: 2014-10-14 – 2014-10-16
• [Presentation] MEMS技術を基盤とした生体ナノポアセンシング (2014)
  • Author(s): 川野竜司, 神谷厚輝, 大崎寿久,竹内昌治
  • Organizer: 日本化学会第94春季年会(2014)
  • Place of Presentation: 名古屋大学 東山キャンパス
  • Year and Date: 2014-03-27 – 2014-03-30
• [Presentation] MICRO-SCALE DROPLET CONTACT METHOD BY MECHANICAL MOTION: REPRODUCIBLE AND ROBUST LIPID BILAYER FORMATION (2013)
  • Author(s): L.N.S. Zaleha, R. Kawano, H. Yasuga, K. Kamiya,T. Osaki, N. Miki and S. Takeuchi
  • Organizer: The Seventeenth International Conference on Miniaturized Systems for Chemistry and Life Sciences
  • Place of Presentation: Freiburg, GERMANY
  • Year and Date: 2013-10-27 – 2013-10-30
• [Book] ナノポア計測：膜タンパク質ナノポアを用いた電気化学的一分子認識 (2014)
  • Author(s): 川野竜司
  • Total Pages: 6
  • Publisher: Chemical Sensors
• [Patent(Industrial Property Rights)] 脂質二重膜形成器具 (2013)
  • Inventor(s): 川野竜司、矢菅浩紀、大崎寿久、神谷厚輝、三木則尚、竹内昌治
  • Industrial Property Rights Holder: KAST、東大
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2013-216009
  • Filing Date: 2013-10-17