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Alteration of antigen recognition and structural dynamics of antibodies during affinity maturation

Research Project

Project/Area Number 18K06161
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43040:Biophysics-related
Research InstitutionKyoto Prefectural University

Principal Investigator

Oda Masayuki  京都府立大学, 生命環境科学研究科, 教授 (20318231)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000)
Fiscal Year 2019: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords抗体 / 抗原結合 / 安定性 / 結晶構造 / 分子認識
Outline of Final Research Achievements

Single-chain Fv (scFv) antibodies against nitrophenyl (NP) on the process of affinity maturation were generated, and their structural, functional, and physical properties were analyzed. The X-ray crystal structure of NP complexed with C6 scFv, affinity-matured type, was determined, showing the structural basis of molecular recognition. The change in thermal stability was analyzed for the scFvs possessing Gly at residue 95 of heavy chain. The stability of germline-type scFv was higher than that of affinity-matured type, which was increased upon NP binding.

Academic Significance and Societal Importance of the Research Achievements

生体内での抗体の親和性成熟の構造基盤を解明し、如何に抗原結合能を高めるかといった抗体医薬分野へも応用可能な知見を得た。また抗体の抗原結合能と熱安定性がトレードオフ関係にあることを示し、蛋白質科学的な抗体の理解のみならず、免疫学的な意義、すなわち抗原結合前の不安定化は結合後の安定化で補われるという知見を得て、今後の蛋白質ラショナルデザイン全般に活かされる。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (10 results)

All 2020 2019 2018 Other

All Journal Article (2 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 2 results,  Open Access: 1 results) Presentation (6 results) (of which Int'l Joint Research: 1 results) Remarks (2 results)

  • [Journal Article] Three-dimensional structure of a high affinity anti-(4-hydroxy-3-nitrophenyl)acetyl antibody possessing a glycine residue at position 95 of the heavy chain2019

    • Author(s)
      Nishiguchi Akihiro、Numoto Nobutaka、Ito Nobutoshi、Azuma Takachika、Oda Masayuki
    • Journal Title

      Molecular Immunology

      Volume: 114 Pages: 545-552

    • DOI

      10.1016/j.molimm.2019.09.001

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Structural and functional evaluation of single-chain Fv antibody HyC1 recognizing the residual native structure of hen egg lysozyme2019

    • Author(s)
      Yamaoka Takanori、Kamatari Yuji O.、Maruno Takahiro、Kobayashi Yuji、Oda Masayuki
    • Journal Title

      Bioscience, Biotechnology, and Biochemistry

      Volume: 84 Issue: 2 Pages: 358-364

    • DOI

      10.1080/09168451.2019.1683441

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] 複数のエピトープを認識する抗体G2の一本鎖Fv抗体を用いた安定性と抗原結合能解析2020

    • Author(s)
      林 隆宏、鎌足 雄司、織田 昌幸
    • Organizer
      第93回 日本生化学会大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 抗体の親和性成熟における抗原結合能と熱安定性の変化2020

    • Author(s)
      西口 明宏、東 隆親、織田 昌幸
    • Organizer
      第56回 熱測定討論会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 抗ニトロフェニル抗体の親和性成熟過程における抗原認識変化の構造基盤2019

    • Author(s)
      西口 明宏、沼本 修孝、伊藤 暢聡、東 隆親、織田 昌幸
    • Organizer
      日本農芸化学会関西支部第508回講演会
    • Related Report
      2019 Research-status Report
  • [Presentation] Increased antigen binding affinity and decreased thermal stability of an anti-(4-hydroxy-3-nitrophenyl)acetyl antibody possessing a glycine residue at position 95 of the heavy chain2019

    • Author(s)
      Masayuki Oda, Takachika Azuma
    • Organizer
      The 33th Annual Symposium of The Protein Society
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Presentation] 抗ニトロフェニル抗体の親和性成熟型一本鎖Fv抗体の抗原認識機構2019

    • Author(s)
      西口 明宏、沼本 修孝、伊藤 暢聡、東 隆親、織田 昌幸
    • Organizer
      第92回日本生化学会大会
    • Related Report
      2019 Research-status Report
  • [Presentation] 抗体の親和性成熟に伴う不安定化と抗原結合に伴う安定化2018

    • Author(s)
      西口 明宏、佐藤 優穂、山岡 敬典、東 隆親、織田 昌幸
    • Organizer
      第91回 日本生化学会大会
    • Related Report
      2018 Research-status Report
  • [Remarks]

    • URL

      https://www2.kpu.ac.jp/life_environ/biophys_chem/

    • Related Report
      2020 Annual Research Report
  • [Remarks]

    • URL

      https://www2.kpu.ac.jp/life_environ/biophys_chem/oda-ref.html

    • Related Report
      2019 Research-status Report

URL: 

Published: 2018-04-23   Modified: 2022-01-27  

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