• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Development of antibodies against extracellular region of receptors by using engineered membrane proteins

Research Project

Project/Area Number 16K01915
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Biomolecular chemistry
Research InstitutionEhime University

Principal Investigator

Takeda Hiroyuki  愛媛大学, プロテオサイエンスセンター, 准教授 (40609393)

Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
KeywordsGPCR / 膜タンパク質 / 抗体 / 受容体 / 人工デザイン抗原 / HRH1 / 無細胞タンパク質合成 / IgG / 膜貫通ドメイン / ヒスタミン受容体 / 抗体医薬 / 細胞外ループ / 人工タンパク質デザイン / 抗原
Outline of Final Research Achievements

Antibodies recognizing extracellular region (ECR) of membrane proteins are important tool required for basic research, structural biology, and development of antibody therapeutics. However, it is very difficult to develop ECR-recognizing antibodies. We hypothesized concealing intracellular region from immune system of host animal, or exposing extracellular region to immune system is effective for induction of ECR-recognizing antibodies.
We selected histamine receptor H1, a class A GPCR, and designed several artificial designed antigen membrane proteins. We produced designed antigens using a cell-free membrane production method. Immunization of those designed antigens to mice successfully induced antibodies recognizing extracellular region of HRH1.

Academic Significance and Societal Importance of the Research Achievements

膜タンパク質は主要な創薬標的である。膜タンパク質の細胞外領域を認識する抗体はフローサイトメトリー等で細胞を識別するマーカーとして欠かせないほか、膜タンパク質の機能構造解析や、抗体医薬開発の点からも求められている。しかし、膜タンパク質の細胞外認識抗体の作製は非常に困難であった。本研究は人工的に改変した膜タンパク質を免疫抗原として用いることで、膜タンパク質の細胞外認識抗体を効率的に取得する方法論を開発することを目的としている。本手法が完成すれば従来よりも高確率、短期間に細胞外認識抗体を開発する事が可能となり、基礎研究から診断薬、医薬開発、バイオセンサーなどの領域に貢献すると期待される。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (16 results)

All 2018 2017 Other

All Journal Article (5 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 4 results,  Open Access: 4 results) Presentation (8 results) (of which Int'l Joint Research: 1 results) Remarks (3 results)

  • [Journal Article] Engineered membrane protein antigens successfully induce antibodies against extracellular regions of claudin-5.2018

    • Author(s)
      Hashimoto Y, et al.
    • Journal Title

      Sci Rep.

      Volume: 8 Issue: 1 Pages: 8383-8383

    • DOI

      10.1038/s41598-018-26560-9

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Functional G-Protein-Coupled Receptor (GPCR) Synthesis: The Pharmacological Analysis of Human Histamine H1 Receptor (HRH1) Synthesized by a Wheat Germ Cell-Free Protein Synthesis System Combined with Asolectin Glycerosomes2018

    • Author(s)
      Suzuki Yasuyuki、Ogasawara Tomio、Tanaka Yuki、Takeda Hiroyuki、Sawasaki Tatsuya、Mogi Masaki、Liu Shuang、Maeyama Kazutaka
    • Journal Title

      Frontiers in Pharmacology

      Volume: 9 Pages: 38-38

    • DOI

      10.3389/fphar.2018.00038

    • Related Report
      2018 Annual Research Report 2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Production of Immunizing Antigen Proteoliposome Using Cell-Free Protein Synthesis System2018

    • Author(s)
      Zhou Wei、Takeda Hiroyuki
    • Journal Title

      Methods and Protocols, Methods in Molecular Biology

      Volume: 1868 Pages: 49-67

    • DOI

      10.1007/978-1-4939-8802-0_7

    • ISBN
      9781493988013, 9781493988020
    • Related Report
      2018 Annual Research Report
  • [Journal Article] Efficiency and Safety of CRAC Inhibitors in Human Rheumatoid Arthritis Xenograft Models2017

    • Author(s)
      Liu Shuang、Hasegawa Hitoshi、Takemasa Erika、Suzuki Yasuyuki、Oka Keizou、Kiyoi Takeshi、Takeda Hiroyuki、Ogasawara Tomio、Sawasaki Tatsuya、Yasukawa Masaki、Maeyama Kazutaka
    • Journal Title

      The Journal of Immunology

      Volume: 199 Issue: 5 Pages: 1584-1595

    • DOI

      10.4049/jimmunol.1700192

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] CP5 system, for simple and highly efficient protein purification with a C-terminal designed mini tag2017

    • Author(s)
      Takeda Hiroyuki、Zhou Wei、Kido Kohki、Suno Ryoji、Iwasaki Takahiro、Kobayashi Takuya、Sawasaki Tatsuya
    • Journal Title

      PLOS ONE

      Volume: 12 Issue: 5 Pages: e0178246-e0178246

    • DOI

      10.1371/journal.pone.0178246

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] GPCRの細胞外領域認識抗体の効率的作製を目指した人工抗原デザイン2018

    • Author(s)
      浜内 孝太郎、澤崎 達也、竹田 浩之
    • Organizer
      第41回分子生物学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 人工デザイン膜タンパク質抗原を用いたClaudin-5細胞領域結合抗体の開発2018

    • Author(s)
      橋本洋祐、Wei Zhou、浜内孝太郎、白倉圭佑、土居健史、八木清仁、澤崎達也、岡田欣晃、近藤昌夫、竹田浩之
    • Organizer
      第41回分子生物学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 人工デザイン膜タンパク質抗原を用いたClaudin-5細胞外領域結合抗体の開発2018

    • Author(s)
      橋本 洋佑、Zhou Wei、浜内 孝太郎、白倉 圭佑、土井 健史、八木 清仁、澤崎 達也、岡田 欣晃、近藤 昌夫、竹田 浩之
    • Organizer
      第13回無細胞生命科学研究会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Cell-free Synthesized Proteoliposome; a New Tool for Antibody Development against Integral Membrane Proteins.2018

    • Author(s)
      Hiroyuki Takeda
    • Organizer
      World Gene Congress 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Accelerate drug discovery by cell-free technology2018

    • Author(s)
      Hiroyuki Takeda
    • Organizer
      Protein Island Matsuyama (PIM) 2018 International Symposium
    • Related Report
      2018 Annual Research Report
  • [Presentation] GPCRの細胞外領域認識抗体の効率的作製を目指した人工抗原デザイン2017

    • Author(s)
      浜内孝太郎、竹田浩之、澤崎達也
    • Organizer
      第17回日本蛋白質科学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] CP5システム:C末ミニタグを用いた高効率タンパク質精製法2017

    • Author(s)
      竹田浩之、城戸康希、Wei Zhou, 矢野智也、寿野良二、小林拓也、澤崎達也
    • Organizer
      第17回日本蛋白質科学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] CP5システム:C末端ミニタグとウサギモノクローナル抗体を用いた高効率タンパク質精製システム2017

    • Author(s)
      竹田浩之、Wei Zhou、城戸康希、寿野良二、岩崎隆弘、小林拓也、澤崎達也
    • Organizer
      ConBio2017(2017年度 生命科学系学会合同年次大会)
    • Related Report
      2017 Research-status Report
  • [Remarks] 愛媛大学プロテオサイエンスセンター プロテオ創薬科学部門 Publications

    • URL

      http://www.pros.ehime-u.ac.jp/proteodrugdiscovery/data/pg23.html

    • Related Report
      2018 Annual Research Report
  • [Remarks] 愛媛大学プロテオサイエンスセンター 研究業績

    • URL

      http://www.pros.ehime-u.ac.jp/result/index.php

    • Related Report
      2018 Annual Research Report
  • [Remarks] 膜タンパク質の細胞外領域に結合する抗体を作るには

    • URL

      http://www.pros.ehime-u.ac.jp/proteodrugdiscovery/Project2/pg22.html

    • Related Report
      2017 Research-status Report

URL: 

Published: 2016-04-21   Modified: 2020-03-30  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi