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Development of a system for stable expression of antibodies bearing non-natural amino acids at defined sites

Research Project

Project/Area Number 25670023
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Biological pharmacy
Research InstitutionOsaka University

Principal Investigator

HINO Nobumasa  大阪大学, 薬学研究科(研究院), 助教 (90469916)

Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywords拡張遺伝暗号 / 非天然型アミノ酸 / tRNA / 抗体薬物複合体 / 非天然アミノ酸 / 部位特異的修飾
Outline of Final Research Achievements

For stable production of antibodies with non-natural amino acids at specific positions, it is necessary to establish cell lines that constitutively and efficiently express exogenous amber suppressor tRNAs. To improve expression efficiency of the exogenous tRNAs, I attempted to develop a system that enables to express them from the nucleolus where cellular tRNAs are transcribed. I constructed tRNA expression vectors containing 5S rDNA sequences or genomic sequences surrounding cellular tRNA genes, which are expected to localize themselves to nucleolus. These vectors successfully increased the expression of the amber mutant of GFP by 1.5- to 2-fold as compared with the vector that contains tRNA genes alone.

Report

(3 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • Research Products

    (5 results)

All 2014 Other

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Presentation (3 results) Remarks (1 results)

  • [Journal Article] Endothelial cell-specific expression of roundabout 4 is regulated by differential DNA methylation of the proximal promoter2014

    • Author(s)
      Okada, Y., Funahashi, N., Tanaka, T., Nishiyama, Y., Yuan, L., Shirakura, K., Turjman, A. S., Kano, Y., Naruse, H., Suzuki, A., Sakai, M., Zhixia, J., Kitajima, K., Ishimoto, K., Hino, N., Kondoh, M., Mukai, Y., Nakagawa, S., Garcia-Cardena, G., Aird, W. C., Doi, T.
    • Journal Title

      Arterioscler Thromb. Vasc. Biol.

      Volume: 34 Issue: 7 Pages: 1531-1538

    • DOI

      10.1161/atvbaha.114.303818

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Presentation] タンパク質間の相互作用部位の解析を可能にする細胞内光クロスリンク技術の開発2014

    • Author(s)
      一瓢奨
    • Organizer
      第13回次世代を担う若手ファーマ・バイオフォーラム2014
    • Place of Presentation
      富山
    • Year and Date
      2014-09-20 – 2014-09-21
    • Related Report
      2014 Annual Research Report
  • [Presentation] 非天然型アミノ酸含有タンパク質の安定生産系の構築2014

    • Author(s)
      喜多絢海、一瓢奨、樋野展正、土井健史
    • Organizer
      日本ケミカルバイオロジー学会 第9回年会
    • Place of Presentation
      大阪
    • Year and Date
      2014-06-11 – 2014-06-13
    • Related Report
      2014 Annual Research Report
  • [Presentation] プローブ転移能を有する光クロスリンカーを用いた細胞内タンパク質間相互作用解析法の開発2014

    • Author(s)
      一瓢奨、喜多絢海、張功幸、高島成二、樋野展正、土井健史
    • Organizer
      日本ケミカルバイオロジー学会 第9回年会
    • Place of Presentation
      大阪
    • Year and Date
      2014-06-11 – 2014-06-13
    • Related Report
      2014 Annual Research Report
  • [Remarks]

    • URL

      http://www.phs.osaka-u.ac.jp/homepage/b018/index.html

    • Related Report
      2014 Annual Research Report

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Published: 2014-07-25   Modified: 2019-07-29  

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