Construction of a system to attain affinity maturation of anti-membrane protein antibodies

• Project/Area Number: 17K19007
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Chemical engineering and related fields
• Research Institution: The University of Tokyo
• Principal Investigator: Kawahara Masahiro 東京大学, 大学院工学系研究科(工学部), 准教授 (50345097)
• Project Period (FY): 2017-06-30 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: シグナル伝達 / 抗体 / 膜蛋白質 / スクリーニング / 親和性成熟

Outline of Final Research Achievements

Membrane proteins are important targets in medicine. Thus, it is necessary to develop a system to obtain antibodies with high specificity and high affinity against membrane proteins in rapid, easy, and reliable ways. In this study we developed a system to rapidly and easily select specific antibodies against a target membrane protein antigen that is displayed on the mammalian cell surface. This system also allowed easy acquisition of antibodies with higher affinity against a target antigen.

Academic Significance and Societal Importance of the Research Achievements

本研究では、既往の抗膜蛋白質抗体選択法の問題点を克服した上で、抗体の親和性成熟を合理的に達成可能なユニークな系の開発に成功しており、学術的に意義深い。本研究の手法を用いて、種々の病態関連膜蛋白質に対する抗体が将来的に取得できれば、抗体医薬品としての臨床応用も含めた大きな社会的波及成果が期待できる。

Research Products

• [Journal Article] An epitope‐directed antibody affinity maturation system utilizing mammalian cell survival as readout (2019)
  • Author(s): Eguchi Akihiro、Nakakido Makoto、Nagatoishi Satoru、Kuroda Daisuke、Tsumoto Kouhei、Nagamune Teruyuki、Kawahara Masahiro
  • Journal Title: Biotechnology and Bioengineering Volume: 116 Issue: 7 Pages: 1742-1751
  • DOI: 10.1002/bit.26965
  • Peer Reviewed
• [Journal Article] Designing Motif-Engineered Receptors To Elucidate Signaling Molecules Important for Proliferation of Hematopoietic Stem Cells. (2018)
  • Author(s): Ishizuka S, Lai CY, Otsu M, Nakauchi H, Nagamune T, Kawahara M.
  • Journal Title: ACS Synth Biol. Volume: 7 Issue: 7 Pages: 1709-1714
  • DOI: 10.1021/acssynbio.8b00163
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A Suicide Switch Directly Eliminates Intracellular scFv Oligomers in the Cytoplasm of Mammalian Cells (2018)
  • Author(s): Nguyen Thuy Duong、Nagamune Teruyuki、Kawahara Masahiro
  • Journal Title: Biotechnology Journal Volume: 14 Issue: 1 Pages: 1800350-1800350
  • DOI: 10.1002/biot.201800350
  • Peer Reviewed
• [Journal Article] 細胞応答シグナルの自在な制御を可能にする受容体改変技術 (2018)
  • Author(s): 河原正浩
  • Journal Title: 化学工学 Volume: 82 Pages: 287-287
• [Journal Article] Dissection of Signaling Events Downstream of the c-Mpl Receptor in Murine Hematopoietic Stem Cells Via Motif-Engineered Chimeric Receptors (2018)
  • Author(s): Saka Koichiro、Lai Chen-Yi、Nojima Masanori、Kawahara Masahiro、Otsu Makoto、Nakauchi Hiromitsu、Nagamune Teruyuki
  • Journal Title: Stem Cell Rev. Rep. Volume: 14 Issue: 1 Pages: 101-109
  • DOI: 10.1007/s12015-017-9768-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Microfluidic preparation of anchored cell membrane sheets for in vitro analyses and manipulation of the cytoplasmic face (2017)
  • Author(s): Izuta,S., Yamaguchi,S., Misawa,R., Yamahira,S., Tan,M., Kawahara,M., Suzuki,T., Takagi,T., Sato,K., Nakamura,M., Nagamune,T. and Okamoto,A.
  • Journal Title: Sci. Rep. Volume: 7:14962 Issue: 1 Pages: 14962-14962
  • DOI: 10.1038/s41598-017-14737-7
  • Peer Reviewed / Open Access
• [Presentation] モチーフ改変受容体を用いた造血幹細胞増殖シグナルの解明 (2019)
  • Author(s): 河原 正浩、石塚周太、坂晃一郎
  • Organizer: 化学工学会第84年会
• [Presentation] 受容体モチーフライブラリのスクリーニングによる効率的な細胞増殖誘導の実現 (2018)
  • Author(s): 梅根 輝来人, 河原 正浩
  • Organizer: 第70回日本生物工学会大会
• [Presentation] 光感受性受容体を利用した標的シグナル伝達分子の光応答的活性化 (2018)
  • 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: 第91回日本生化学会大会
• [Presentation] A receptor-engineering approach for arbitrarily controlling cell fates (2018)
  • Author(s): Masahiro Kawahara
  • Organizer: The University of Tokyo-Tsinghua University Joint Symposium 2018
  • Int'l Joint Research / Invited
• [Presentation] ゲノム編集ツールを用いた細胞運命変換系の構築 (2018)
  • Author(s): 河原 正浩
  • Organizer: 化学工学会第50回秋季大会
• [Presentation] 受容体ライブラリーによる機能性細胞の創製 (2018)
  • Author(s): 河原 正浩
  • Organizer: 第70回日本生物工学会大会
  • Invited
• [Presentation] 細胞内シグナル伝達の合成生物工学 (2017)
  • Author(s): 河原 正浩
  • Organizer: 第7 回合成生物工学シンポジウム
  • Invited
• [Presentation] Development of an Epitope selective Antibody Selection System Using a Cell Survival as a Readout (2017)
  • Author(s): Akihiro Eguchi, Teruyuki Nagamune, Masahiro Kawahara
  • Organizer: YABEC2017
  • Int'l Joint Research
• [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 (ConBio2017)
• [Presentation] モチーフのフェノタイプスクリーニングによる細胞運命制御受容体の効率的な開発 (2017)
  • Author(s): 梅根 輝来人、長棟 輝行、河原 正浩
  • Organizer: 生命科学系学会合同年次大会 (ConBio2017)
• [Presentation] 細胞生存を指標としたエピトープ選択的抗体選択法の開発 (2017)
  • Author(s): 江口 晃弘、長棟 輝行、河原 正浩
  • Organizer: 第69回日本生物工学会大会
• [Presentation] 効率的な細胞運命制御を指向したチロシンモチーフのスクリーニング (2017)
  • Author(s): 梅根 輝来人、長棟 輝行、河原 正浩
  • Organizer: 第69回日本生物工学会大会
• [Book] Biomedical Engineering Challenges: A Chemical Engineering Insight (2018)
  • Author(s): Kawahara, M.
  • Total Pages: 26
  • Publisher: Wiley
  • ISBN: 9781119296034