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Application of FALI with antibody-like scaffolds to the high-throughput functional proteomic screen for therapeutic targets

Research Project

Project/Area Number 26670127
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field General pharmacology
Research InstitutionJuntendo University

Principal Investigator

SAKURAI Takashi  順天堂大学, 医学部, 教授 (70225845)

Co-Investigator(Kenkyū-buntansha) NEMOTO Naoto  埼玉大学, 理工学研究科, 教授 (60509727)
Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywords神経科学 / プロテオーム / 組換え抗体 / 薬理学
Outline of Final Research Achievements

Fluorophore-assisted light inactivation (FALI) in combination with a cell-based assay is a unique method for functional proteomic screen to search for regulators of a specific cellular function. One of the major obstacles for this method is the construction of antibody libraries containing high-affinity binders that cover cell surface membrane proteins as potential targets. To overcome barriers to efficiency, we tested new scaffolds compatible with cDNA display method. By using a single domain antibody framework derived from camel heavy-chain antibodies (VHH or nanobody), we optimized the cell-free expression of recombinant antibody clones that are linked to their genotype. Now we are trying pilot-scale clone selection for erbB as a model system.

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • Research Products

    (5 results)

All 2015 2014 Other

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 2 results,  Acknowledgement Compliant: 2 results) Presentation (2 results) Remarks (1 results)

  • [Journal Article] A versatile puromycin-linker using cnvK for high-throughput in vitro selection by cDNA display2015

    • Author(s)
      Yuki Mochizuki, Takeru Suzuki, Kenzo Fujimoto, Naoto Nemoto
    • Journal Title

      Journal of Biotechnology

      Volume: 212 Pages: 174-180

    • DOI

      10.1016/j.jbiotec.2015.08.020

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Easy and rapid binding assay for functional analysis of disulfide-containing peptides by a pull-down method using a puromycin-linker and a cell-free translation system2015

    • Author(s)
      Yutaro Tanemura, Yuki Mochizuki, Shigefumi Kumachi and Naoto Nemoto
    • Journal Title

      Biology

      Volume: 4 Issue: 1 Pages: 161-172

    • DOI

      10.3390/biology4010161

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] 試験管内選択により取得されたジスルフィドリッチペプチドアプタマーは分子認識スキャフォールドとなりうるか?2014

    • Author(s)
      望月 佑樹, 根本 直人
    • Organizer
      第37回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県・横浜市)
    • Year and Date
      2014-11-26
    • Related Report
      2014 Research-status Report
  • [Presentation] 人工合成ラクダ科単ドメイン抗体(VHH)ライブラリを用いたsurvivin結合分子のcDNA display法による試験管内淘汰2014

    • Author(s)
      木村 真之介, 望月 佑樹, 鈴木 武尊, 根本 直人
    • Organizer
      第37回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県・横浜市)
    • Year and Date
      2014-11-25
    • Related Report
      2014 Research-status Report
  • [Remarks] 組換え抗体ライブラリーと光による蛋白質機能不活性化法(FALI)を用いた創薬標的探索

    • URL

      http://pharmacology.sakura.ne.jp/jp/research/fali_res/fali_res.html

    • Related Report
      2015 Annual Research Report 2014 Research-status Report

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Published: 2014-04-04   Modified: 2017-05-10  

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