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2020 年度 実績報告書

Understanding the efficacy of therapeutic antibodies through their interaction with cellular receptors.

研究課題

研究課題/領域番号 20H03228
研究機関九州大学

研究代表者

CAAVEIRO Jose  九州大学, 薬学研究院, 教授 (00536732)

研究分担者 高橋 大輔  九州大学, 薬学研究院, 講師 (70791523)
研究期間 (年度) 2020-04-01 – 2023-03-31
キーワードantibody / fc receptor / hinge / structure / immunology / glycosylation / protein expression
研究実績の概要

Key events contributing to the efficacy of therapeutic antibodies, such as the activation of cellular Fcgamma receptors, are still poorly understood. The purpose of this research is to understand the molecular mechanism of antibodies and their interaction with their cellular Fc gamma receptors.
We are applying biophysical and structural techniques to understand it. The constructs (antibodies, Fc receptors, vectors), instruments (such a new purification system and C)2 incubators), and many necessary protocols for protein preparation have been set-up. The expression of some recombinant proteins has been also achieved.
Antibodies with greater affinity for their cognate antigens are highly desirable to achieve the goals of this project. We have developed a new concept resulting in a new modality antibody with greater affinity using site-directed chemical modification. This work resulted in a publication showing increase of potency of up to 700-fold against the target.
A new approach to study the Fc region is also underway, consisting in the simultaneous production of Fc region in the glycosylated as well as in the un-glycosylated fashion. We have been able to obtain recombinant protein in high-yield, and the results have been published.
Finally, attempts to crystallize IgG1 and Fc receptors is also underway.

現在までの達成度 (区分)
現在までの達成度 (区分)

2: おおむね順調に進展している

理由

Despite the tough working environment at universities in Japan, the project has advanced adequately as demonstrated by the two publications achieved. In addition, we have been able to overcome numerous hurdles to establish a smooth workflow to steadily advance the project. Unfortunately because of the tough circumstances derived from the pandemics I was not able to attend international conferences.

今後の研究の推進方策

Basically to continue with the research lines started this year.
(1) preparation of specimens for structural analysis by X-ray crystallography and if possible Cryo-EM
(2) Understand the contribution of glycosylation and other modification in the properties of antibodies
(3) Evaluate the role of the hinge region by mutational analysis
(4) Attempt to quantify the clustering effect in model systems using the membrane portion of the receptor.

  • 研究成果

    (2件)

すべて 2021 2020

すべて 雑誌論文 (2件) (うち国際共著 2件、 査読あり 2件、 オープンアクセス 1件)

  • [雑誌論文] High-level expression of human CH2 domain from the Fc region in <i>Pichia pastoris</i> and preparation of anti-CH2 antibodies2021

    • 著者名/発表者名
      Oyama Kosuke、Ohkuri Takatoshi、Inoue Mao、Caaveiro Jose M M、Ueda Tadashi
    • 雑誌名

      The Journal of Biochemistry

      巻: 170 ページ: 289~297

    • DOI

      10.1093/jb/mvab039

    • 査読あり / 国際共著
  • [雑誌論文] Affinity for the Interface Underpins Potency of Antibodies Operating In Membrane Environments2020

    • 著者名/発表者名
      Rujas Edurne、various authors, Ojida Akio、Domene Carmen、Caaveiro Jose M.M.、Nieva Jose L.
    • 雑誌名

      Cell Reports

      巻: 32 ページ: 108037~108037

    • DOI

      10.1016/j.celrep.2020.108037

    • 査読あり / オープンアクセス / 国際共著

URL: 

公開日: 2022-12-28  

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