Simultaneous detection system for multiple disease markers in blood using aptamer set

• Project/Area Number: 24360338
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: IKEBUKURO kazunori 東京農工大学, 工学(系)研究科(研究院), 教授 (70251494)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥19,240,000 (Direct Cost: ¥14,800,000、Indirect Cost: ¥4,440,000); Fiscal Year 2014: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000); Fiscal Year 2013: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2012: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
• Keywords: アプタマー / 疾病マーカー / 同時検出 / バイオ関連機器 / 診断技術 / 高感度検出 / 次世代シーケンサー / 固定化ビーズ / 血液測定 / PCR

Outline of Final Research Achievements

We developed the method to simultaneously detect multiple disease markers in blood using a set of aptamers against various disease markers in blood. We immobilized the various markers in blood onto the beads and added a set of aptamers onto that. We then extracted the bound aptamers from the beads and determined those sequences. We can estimate the concentration of the specific disease marker by counting the sequences of its specific aptamer.
We checked our proposed detection system worked but the background signal from the unspecific binding of the aptamers to the beads or other proteins apart from the target protein is so high that it seemed hard to realize the highly sensitive detection at this time. We found the advantage of the use of boronic acid immobilized bead, which does not seem to affect the structure of proteins upon immobilization. That would become important tool for development of the detection system using aptamers.

Research Products

• [Journal Article] Selection of DNA aptamers against uropathogenic Escherichia coli NSM59 by quantitative PCR controlled Cell-SELEX (2014)
  • Author(s): Nasa Savory, Jonathan Nzakizwanayo, Koichi Abe, Wataru Yoshida, Stefano Ferri, Cinzia Dedi, Brian V Jones, Kazunori Ikebukuro
  • Journal Title: Journal of Microbiological Methods Volume: 104 Pages: 94-100
  • DOI: 10.1016/j.mimet.2014.06.016
  • Peer Reviewed
• [Journal Article] Emerging techniques employed in aptamer-based diagnostic tests (2014)
  • Author(s): Yoshida W, Abe K, Ikebukuro K
  • Journal Title: Expert Rev Mol Diagn Volume: 14 Issue: 2 Pages: 143-151
  • DOI: 10.1586/14737159.2014.868307
  • Peer Reviewed
• [Journal Article] Electrochemical biosensors using aptamers for theranostics (2014)
  • Author(s): Abe K, Yoshida W, Ikebukuro K
  • Journal Title: Adv Biochem Eng Biotechnol Volume: 140 Pages: 183-202
  • Peer Reviewed
• [Presentation] In silico maturation: Advanced aptamer development for biosensing applications (2015)
  • Author(s): Savory N., Jones B., Abe K., Ikebukuro K.
  • Organizer: Biosensor2014
  • Place of Presentation: オーストラリア、メルボルン
  • Year and Date: 2015-05-27 – 2015-05-30
• [Presentation] Screening and improvement of DNA aptamers by in silico approaches (2015)
  • Author(s): Yokoyama T., Yoshida W.,Saito T., Ikebukuro K.
  • Organizer: Biosensor2014
  • Place of Presentation: オーストラリア、メルボルン
  • Year and Date: 2015-05-27 – 2015-05-30
• [Presentation] in silico maturation 法を用いたアプタマーの機能改良 (2015)
  • Author(s): 池袋一典
  • Organizer: 第１０回理研「バイオものづくり」シンポジウム
  • Place of Presentation: 和光市 理化学研究所
  • Year and Date: 2015-03-06
• [Presentation] In silico maturation: improvement of aptamer function based on genetic algorithms employing in vitro functional screening process (2014)
  • Author(s): Nasa S., Yokoyama T., Saito T., Nonaka Y, Fukaya T., Yoshida W., Abe K., Ikebukuro K.
  • Organizer: The 41th International Symposium on Nucleic Acids Chemistry
  • Place of Presentation: 北九州市、甲南大学
  • Year and Date: 2014-11-05 – 2014-11-07
• [Book] 高分子10月号「核酸リガンド“アプタマー”のリプログラミング」 (2014)
  • Author(s): 池袋一典、セーボレー那沙、阿部公一
  • Total Pages: 4
  • Publisher: 公益社団法人高分子学会
• [Book] "In silico Maturation: Processing Sequences to Improve Biopolymer Functions Based on Genetic Algorithms" in "Applications of Metaheuristics in Process Engineering" (2014)
  • Author(s): Savory, N., Abe, K., Yoshida, W., & Ikebukuro, K
  • Publisher: Springer