Development of sensitive and inexpensive antibody array chip

• Project/Area Number: 24760649
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biofunction/Bioprocess
• Research Institution: Kyoto Institute of Technology
• Principal Investigator: KUMADA Yoichi 京都工芸繊維大学, 工芸科学研究科, 准教授 (70452373)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2012: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: PMMA-tag / 単鎖抗体 / 配向固定 / マイクロアレイ / 固定化 / イムノアッセイ

Research Abstract

In this study, we investigated to establish technologies for fabricatioin of antibody-based microarray chips immobilized with single-chain Fv antibodies (scFvs) by utilizing our original solid-phase refolding technology and conventional microarray spotting technology. Consequently, we found that our originally-isolated peptide tag specific to poly(methyl methacrylate) was significantly useful for efficient refolding and site-specific immobilization of scFv onto the surface of PMMA plate. Thus, we successfully established preparation method of scFv-immobilized PMMA plate and sensitive detection of antigen could be attained.

Research Products

• [Journal Article] Immobilization and functional reconstitution of antibody Fab fragment by solid-phase refolding (2014)
  • Author(s): Y. Kumada, K. Hamasaki, A. Nakagawa, E. Sasaki, T. Shirai, M. Okumura, M. Inoue, M. Kishimoto
  • Journal Title: Journal of Immunological Methods
• [Journal Article] Identification and characterization of peptide fragments for the direct and site-specific immobilization of functional proteins onto the surface of silicon nitride (2014)
  • Author(s): Y. Kumada, T. Ootsuka, M. Asada, S. Yoshizuka, M. Chiyama, M. Sakane, F. MD Hasan, K. Sawada, K. Okumura, M. Kishimoto
  • Journal Title: Journal of Biotechnology Volume: in press
  • Peer Reviewed
• [Journal Article] Immobilization and functional reconstitution of antibody Fab fragment by solid-phase refolding (2013)
  • Author(s): Yoichi Kumada, Kyoko Hamasaki, Aya Nakagawa, Eiju Sasaki, Tatsinori Shirai, Masahiro Okumura, Manami Inoue, Michimasa Kishimoto
  • Journal Title: Journal of Immunological Methods Volume: 400 - 401 Pages: 70-77
  • DOI: 10.1016/j.jim.2013.10.006
  • Peer Reviewed
• [Journal Article] Identification and characterization of peptide fragments for the direct and site-specific immobilization of functional proteins onto the surface of silicon nitride
  • Author(s): Y. Kumada, T. Ootsuka, M. Asada, S. Yoshizuka, M. Chiyama, M. Sakane, F. MD Hasan, K. Sawada, K. Okumura, M. Kishimoto
  • Journal Title: Journal of Biotechnology Volume: (in press)
• [Book] 新生物化学工学 (2013)
  • Author(s): 岸本通雅、堀内淳一、藤原伸介、熊田陽一
  • Total Pages: 198
  • Publisher: 三共出版
• [Patent(Industrial Property Rights)] 抗体のリフォールディング方法、リフォールディングされた抗体の製造方法、リフォールディングされた抗体、及びこれらの利用 (2013)
  • Inventor(s): 熊田陽一、岸本通雅、石川泰行、藤原勇佐
  • Industrial Property Rights Holder: 京都工芸繊維大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2013-027862
  • Filing Date: 2013-02-15