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Development of intrabody selection technology in mammalian cells

Research Project

Project/Area Number 16K14486
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Biofunction/Bioprocess
Research InstitutionThe University of Tokyo

Principal Investigator

Nagamune Teruyuki  東京大学, 大学院工学系研究科(工学部), 教授 (20124373)

Co-Investigator(Renkei-kenkyūsha) KAWAHARA Masahiro  東京大学, 大学院工学系研究科, 准教授 (50345097)
Project Period (FY) 2016-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywordsバイオテクノロジー / キメラ受容体 / 細胞内抗体スクリーニング / 細胞内蛋白質間相互作用検出 / 細胞アレイ
Outline of Final Research Achievements

In this study, for the development of next-generation antibody drug, we developed a novel versatile intrabody selection method, which can select intrabody against antigen protein expressed in the cytosol of mammalian cells rapidly and directly. We constructed a chimeric receptor library, which can transduce growth signal by binding to antigen, by conjugating naive library of single chain antibody scFv and intracellular domain of receptor c-kit through flexible linker G4S. We expressed this chimeric receptor library together with a model antigen, the nucleoprotein of a rabies virus, in IL-3 dependent mammalian cell Ba/F3, and successfully obtained several scFv binders against the antigen protein from proliferated cells under IL-3 deficient culture condition.

Report

(3 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • Research Products

    (19 results)

All 2017 2016

All Journal Article (4 results) (of which Peer Reviewed: 4 results,  Open Access: 4 results,  Acknowledgement Compliant: 1 results) Presentation (15 results) (of which Int'l Joint Research: 6 results)

  • [Journal Article] Engineering a growth sensor to select intracellular antibodies in the cytosol of mammalian cells Journal of Bioscience and Bioengineering2017

    • Author(s)
      Nguyen TD, Takasuka H, Kaku Y, Inoue S, Nagamune T, Kawahara M
    • Journal Title

      Journal of Bioscience and Bioengineering

      Volume: 印刷中 Issue: 1 Pages: 125-132

    • DOI

      10.1016/j.jbiosc.2017.02.017

    • Related Report
      2017 Annual Research Report 2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] A Chemically Inducible Helper Module for Detecting Protein-Protein Interactions with Tunable Sensitivity Based on KIPPIS2017

    • Author(s)
      Kashima Daiki、Kawade Raiji、Nagamune Teruyuki、Kawahara Masahiro
    • Journal Title

      Anal. Chem.

      Volume: 89 Issue: 9 Pages: 4824-4830

    • DOI

      10.1021/acs.analchem.6b04063

    • Related Report
      2017 Annual Research Report 2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Ligand-inducible dimeric antibody for selecting antibodies against a membrane protein based on mammalian cell proliferation2016

    • Author(s)
      Miura, T., Nagamune, T., Kawahara, M.
    • Journal Title

      Biotechnol. Bioeng.

      Volume: 113 Issue: 5 Pages: 1113-1123

    • DOI

      10.1002/bit.25858

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Growth signalobody selects functional intrabodies in the mammalian cytoplasm2016

    • Author(s)
      Lee S, Kaku Y, Inoue S, Nagamune T, Kawahara M.
    • Journal Title

      Biotechnol Journal

      Volume: 11 Issue: 4 Pages: 565-573

    • DOI

      10.1002/biot.201500364

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 細胞内タンパク質間相互作用検出系の構築と阻害剤探索への展開2017

    • Author(s)
      鹿島大揮, 河出来時, 河原正浩, 長棟輝行
    • Organizer
      生物工学若手研究者の集い 夏のセミナー
    • Related Report
      2017 Annual Research Report
  • [Presentation] Synthetic Control of Mammalian Cell Signaling by Engineering Receptor Tyrosine Kinase2017

    • Author(s)
      Tatphon Kongrongtong, Teruyuki Nagamune and Masahiro Kawahara
    • Organizer
      The 13th Asian Congress on Biotechnology, Bioinnovation and Bioeconomy 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 受容体の分子改変による細胞内抗体選択法の開発2017

    • Author(s)
      河原正浩, グエン トゥイズオン, 李 松熹, 高須賀 仁, 長棟輝行
    • Organizer
      化学工学会第49回秋季大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] On-targetシグナル伝達分子の特異的活性化を目的としたデザイナー受容体の開発2017

    • Author(s)
      コンクローントーン タットポーン, 長棟輝行, 河原正浩
    • Organizer
      生命科学系学会合同年次キメラ受容体の細胞内局在最適化による高感度タンパク質間相互作用検出系の確立大会(第40回日本分子生物学会年会 第90回日本生化学会大会)
    • Related Report
      2017 Annual Research Report
  • [Presentation] キメラ受容体の細胞内局在最適化による高感度タンパク質間相互作用検出系の確立2017

    • Author(s)
      垣内洋祐, 長棟輝行, 河原正浩
    • Organizer
      生命科学系学会合同年次大会(第40回日本分子生物学会年会 第90回日本生化学会大会)
    • Related Report
      2017 Annual Research Report
  • [Presentation] 細胞増殖を指標とした細胞内タンパク質間相互作用検出系 (KIPPIS) の構築と阻害剤探索への展開2017

    • Author(s)
      鹿島大揮, 河出来時, 長棟輝行, 河原正浩
    • Organizer
      生命科学系学会合同年次大会(第40回日本分子生物学会年会 第90回日本生化学会大会)
    • Related Report
      2017 Annual Research Report
  • [Presentation] 狙った結合部位上における抗体親和性成熟系の開発2017

    • Author(s)
      江口晃弘, 長棟輝行, 河原正浩
    • Organizer
      生命科学系学会合同年次大会(第40回日本分子生物学会年会 第90回日本生化学会大会)
    • Related Report
      2017 Annual Research Report
  • [Presentation] A novel cell-based intracellular protein-protein interaction detection platform (KIPPIS) for intrabody screening2017

    • Author(s)
      Daiki Kashima, Raiji Kawade, Teruyuki Nagamune, Masahiro Kawahara
    • Organizer
      Antibody Engineering & Therapeutics
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Developing a suicide switch for improving quality of scFv libraries2016

    • Author(s)
      Thuy Duong Nguyen, Teruyuki Nagamune, Masahiro Kawahara
    • Organizer
      Antibody Engineering & Therapeutics 2016
    • Place of Presentation
      Manchester Grand Hyatt San Diego (San Diego, U.S.A.)
    • Year and Date
      2016-12-12
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] 細胞内タンパク質間相互作用検出系のホモ二量体化ヘルパーモジュールを利用した高感度化2016

    • Author(s)
      鹿島大揮, 河出来時, 長棟輝行, 河原正浩
    • Organizer
      「細胞を創る」研究会9.0
    • Place of Presentation
      早稲田大学 国際会議場 井深大記念ホール(東京都新宿区)
    • Year and Date
      2016-11-21
    • Related Report
      2016 Research-status Report
  • [Presentation] Highly sensitive detection of intracellular protein‐protein interactions by fusing chemically inducible homo-dimeric module2016

    • Author(s)
      Daiki Kashima, Raiji Kawade, Teruyuki Nagamune, Masahiro Kwahara
    • Organizer
      The 29th Annual and International Meeting of the Japanese Association for Animal Cell Technology
    • Place of Presentation
      神戸国際会議場(兵庫県神戸市)
    • Year and Date
      2016-11-09
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Development of an Epitope-selective Antibody Selection System Using a Cell Death Signal2016

    • Author(s)
      Akihiro Eguchi, Teruyuki Nagamune, Masahiro Kawahara
    • Organizer
      The 29th Annual and International Meeting of the Japanese Association for Animal Cell Technology
    • Place of Presentation
      神戸国際会議場(兵庫県神戸市)
    • Year and Date
      2016-11-09
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] KIPPIS: Probing intracellular protein-protein interactions with mammalian cell growth2016

    • Author(s)
      Masahiro Kawahara, Satoru Mabe, Teruyuki Nagamune
    • Organizer
      The 22nd symposium of Young Asian Biological Engineers' Community
    • Place of Presentation
      フェニックスシーガイヤリゾート(宮崎県宮崎市)
    • Year and Date
      2016-10-27
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] ホモ二量体化モジュールの導入による細胞内タンパク質間相互作用検出の高感度化2016

    • Author(s)
      鹿島大揮, 河出来時, 長棟輝行, 河原正浩
    • Organizer
      第68回日本生物工学会大会
    • Place of Presentation
      富山国際会議場(富山県富山市)
    • Year and Date
      2016-09-28
    • Related Report
      2016 Research-status Report
  • [Presentation] 哺乳類細胞質内を反応場とした細胞内抗体選択プラットホームの開発2016

    • Author(s)
      鹿島大揮, 長棟輝行, 河原正浩
    • Organizer
      2016年度生物工学若手研究者の集い(若手会)夏のセミナー
    • Place of Presentation
      ホテルコンチネンタル府中(東京都府中市)
    • Year and Date
      2016-07-16
    • Related Report
      2016 Research-status Report

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Published: 2016-04-21   Modified: 2019-03-29  

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