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Development of a generalizable toolkit for rapidly improving the sensitivity of fluorescent immunosensor proteins

Research Project

Project/Area Number 21K14468
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 27040:Biofunction and bioprocess engineering-related
Research InstitutionTokyo Institute of Technology

Principal Investigator

Zhu Bo  東京工業大学, 科学技術創成研究院, 助教 (70886605)

Project Period (FY) 2021-04-01 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2022: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2021: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywordsbiosensor / uHTS / quenchbody / mRNA display / phage display / Immunosensor / Quenchbody / SARS-CoV-2 / Phage display / NGS / バイオテクノロジー / タンパク質 / バイオセンサー / 蛍光 / mRNA-display
Outline of Research at the Start

Quenchbody(Q-body)は, 抗体あるいはその断片を部位特異的に蛍光修飾して得られる, 蛍光免疫センサーである. Q-bodyは, クエンチ解消原理に基づき様々な抗原を迅速・簡便に検出可能と考えられている. しかし, 現状では, 一つの抗原が一つのQ-bodyの蛍光色素を光らせる事しかできず, Q-bodyの感度は, 親抗体の動力学的特性によって制限されると考えられる. 本研究では, Q-bodyの感度を短期間で向上させるためのツールキットを開発することを目的とする. このツールキットにより, Q-bodyに基づく免疫測定法の開発期間を短縮し, 緊急事態への迅速な対応を可能にする.

Outline of Final Research Achievements

In this project, a toolkit for rapidly improving the performance of Quenchbody biosensors has been developed. It includes a biomolecular display-based ultra-high-throughput screening system, a cleavable fluorophore linker design for increasing the detection limit of the biosensor, and a reaction buffer with optimized crowding agents. Several Quenchbody biosensors, including the SARS-CoV-2 N protein biosensor, were optimized using this toolkit and rapidly reached a performance level suitable for the detection of analytes in practical applications. This toolkit, especially the uHTS screening method, will change the development process of Quenchbody biosensors or similar antibody conjugates, allowing the creation of biosensors in a timely manner under emergency circumstances in the future.

Academic Significance and Societal Importance of the Research Achievements

バイオセンサーおよび関連する均一系イムノアッセイは、診断や食品安全の分野において、ターゲットバイオ分子の迅速かつ高感度な検出において極めて重要な役割を果たしています。本研究では、Quenchbodyバイオセンサーの迅速な設計と開発のためのツールキットを開発しました。このツールキットは、従来の手法に比べて設計および最適化プロセスを大幅に効率化し、短時間でバイオセンサーを作成することを可能にします。これにより、将来的にパンデミックなどの緊急事態に迅速かつ効果的に対応できる社会の実現に寄与することが期待されます。

Report

(3 results)
  • 2023 Final Research Report ( PDF )
  • 2022 Research-status Report
  • 2021 Research-status Report
  • Research Products

    (22 results)

All 2023 2022 2021 Other

All Int'l Joint Research (1 results) Journal Article (7 results) (of which Int'l Joint Research: 4 results,  Peer Reviewed: 7 results,  Open Access: 5 results) Presentation (12 results) (of which Int'l Joint Research: 2 results) Remarks (2 results)

  • [Int'l Joint Research] Weifang Medical University(中国)

    • Related Report
      2022 Research-status Report
  • [Journal Article] The Patrol Yeast: A new biosensor armed with antibody-receptor chimera detecting a range of toxic substances associated with food poisoning2023

    • Author(s)
      Jiulong Su, Bo Zhu, Akihito Inoue, Hiroyuki Oyama, Izumi Morita, Jinhua Dong, Takanobu Yasuda, Yoshiko Sugita-Konishi, Tetsuya Kitaguchi, Norihiro Kobayashi, Shiro Miyake, Hiroshi Ueda
    • Journal Title

      Biosensors and Bioelectronics

      Volume: 219 Pages: 114793-114793

    • DOI

      10.1016/j.bios.2022.114793

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Recent progress in homogeneous immunosensors based on fluorescence or bioluminescence using antibody engineering2023

    • Author(s)
      Rani Abdul Qawee、Zhu Bo、Ueda Hiroshi、Kitaguchi Tetsuya
    • Journal Title

      Analyst

      Volume: 148 Issue: 7 Pages: 1422-1429

    • DOI

      10.1039/d2an01913b

    • Related Report
      2022 Research-status Report
    • Peer Reviewed
  • [Journal Article] Rapid and sensitive SARS-CoV-2 detection using a homogeneous fluorescent immunosensor Quenchbody with crowding agents2022

    • Author(s)
      Zhu Bo、Nosaka Nobuyuki、Kanamaru Shuji、Dong Jinhua、Dai Yancen、Inoue Akihito、Yang Yinghui、Kobayashi Kaori、Kitaguchi Tetsuya、Iwasaki Hiroshi、Koike Ryuji、Wakabayashi Kenji、Ueda Hiroshi
    • Journal Title

      The Analyst

      Volume: 147 Issue: 22 Pages: 4971-4979

    • DOI

      10.1039/d2an01051h

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Isolation of a human SARS-CoV-2 neutralizing antibody from a synthetic phage library and its conversion to fluorescent biosensors2022

    • Author(s)
      Li Haimei、Zhu Bo、Li Baowei、Chen Limei、Ning Xuerao、Dong Hang、Liang Jingru、Yang Xueying、Dong Jinhua、Ueda Hiroshi
    • Journal Title

      Scientific Reports

      Volume: 12 Issue: 1 Pages: 15496-15496

    • DOI

      10.1038/s41598-022-19699-z

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Creating a Thermostable β-Glucuronidase Switch for Homogeneous Immunoassay by Disruption of Conserved Salt Bridges at Diagonal Interfaces2022

    • Author(s)
      Zhu Bo、Qian Cheng、Tang Haoxuan、Kitaguchi Tetsuya、Ueda Hiroshi
    • Journal Title

      Biochemistry

      Volume: 62 Issue: 2 Pages: 309-317

    • DOI

      10.1021/acs.biochem.2c00165

    • Related Report
      2022 Research-status Report
    • Peer Reviewed
  • [Journal Article] Intra Q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live cell imaging and sorting2022

    • Author(s)
      Dai Yancen、Sato Yuko、Zhu Bo、Kitaguchi Tetsuya、Kimura Hiroshi、Ghadessy Farid J.、Ueda Hiroshi
    • Journal Title

      Chemical Science

      Volume: 13 Issue: 33 Pages: 9739-9748

    • DOI

      10.1039/d2sc02355e

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Evaluation and selection of potent fluorescent immunosensors by combining fluorescent peptide and nanobodies displayed on yeast surface2021

    • Author(s)
      Inoue Akihito、Yasuda Takanobu、Zhu Bo、Kitaguchi Tetsuya、Murakami Akikazu、Ueda Hiroshi
    • Journal Title

      Scientific Reports

      Volume: 11 Issue: 1 Pages: 22590-22590

    • DOI

      10.1038/s41598-021-02022-7

    • NAID

      120007172601

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Rapid construction of homogeneous immunosensors for SARS-CoV-2 nucleocapsid and spike proteins using Quenchbody and Quenchprobe technology2022

    • Author(s)
      Bo Zhu, Haimei Li, Yancen Dai, Akihito Inoue, Yinghui Yang, Kaori Kobayashi, Tetsuya Kitaguchi, Jinhua Dong, Hiroshi Ueda
    • Organizer
      Bioanalytical Sensors Gordon Research Conference
    • Related Report
      2022 Research-status Report
    • Int'l Joint Research
  • [Presentation] Converting antibodies to rapid response SARS-CoV-2 homogeneous immunosensors via Quenchbody technology and crowding agent2022

    • Author(s)
      朱博, 野坂 宜之, 董 金華, 北口 哲也, 小池 竜 司, 若林 健二, 上田 宏
    • Organizer
      化学工学会第53回秋季大会
    • Related Report
      2022 Research-status Report
  • [Presentation] 蛍光免疫センサーQuenchbodyのシグナル増幅と高速スクリーニング法の探索2022

    • Author(s)
      塘 知子, 朱 博, 北口 哲也, 上田 宏
    • Organizer
      第95回日本生化学会大会
    • Related Report
      2022 Research-status Report
  • [Presentation] Intra Q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live-cell imaging and sorting2022

    • Author(s)
      Dai Yancen, Sato Yuko, Zhu Bo, Kitaguchi Tetsuya, Kimura Hiroshi, Farid J. Ghadessy, Ueda Hiroshi
    • Organizer
      Bioanalytical Sensors Gordon Research Conference
    • Related Report
      2022 Research-status Report
    • Int'l Joint Research
  • [Presentation] 3分割ルシフェラーゼを用いた免疫測定法における相互作用の測定と数理モデルの作製2022

    • Author(s)
      佐藤 里杏人, 安田 貴信, 朱 博, 北口 哲也, 加藤 直洋, 上田 宏
    • Organizer
      化学工学会第88年会
    • Related Report
      2022 Research-status Report
  • [Presentation] 酵母細胞表層提示系を用いたナノ抗体由来”nano Q-body”のTrp変異進化2022

    • Author(s)
      井上暁人, 安田貴信, 朱博, 北口哲也, 村上明一, 上田宏
    • Organizer
      化学工学会第88年会
    • Related Report
      2022 Research-status Report
  • [Presentation] コルチゾール認識ナノ抗体の蛍光免疫センサー化2022

    • Author(s)
      井上暁人, 鈴木大凱, YANG Yinghui, 安田貴信, 朱博, 北口哲也, 上田宏
    • Organizer
      第16回バイオ関連化学シンポジウム
    • Related Report
      2022 Research-status Report
  • [Presentation] Quenchbodyを用いた1分間SARS-CoV-2及び変異株の検出法2021

    • Author(s)
      朱 博、董 金華、寧 雪じょう、井上 暁人、北口 哲也、上田 宏
    • Organizer
      化学工学会第52回秋季大会
    • Related Report
      2021 Research-status Report
  • [Presentation] 蛍光免疫プローブQuenchprobeのダブルラベル化の検討2021

    • Author(s)
      小林 香央里、朱 博、北口 哲也、上田 宏
    • Organizer
      化学工学会第87年会
    • Related Report
      2021 Research-status Report
  • [Presentation] 細胞内腫瘍マーカーp53可視化のための免疫蛍光プローブQ-bodyの開発2021

    • Author(s)
      Dai Yancen、朱 博、佐藤 優子、木村 宏、Ghadessy Farid J、北口 哲也、上田 宏
    • Organizer
      化学工学会第87年会
    • Related Report
      2021 Research-status Report
  • [Presentation] 蛍光免疫センサーCoiled Quenchbodyのがん光線力学療法への応用2021

    • Author(s)
      椿 凜果、安田 貴信、朱 博、北口 哲也、上田 宏
    • Organizer
      化学工学会第87年会
    • Related Report
      2021 Research-status Report
  • [Presentation] 培養液中の組換えタンパク質が検出可能な蛍光免疫センサの構築2021

    • Author(s)
      寧 せつじょう、安田 貴信、朱 博、北口 哲也、上田 宏
    • Organizer
      第15回バイオ関連化学シンポジウム
    • Related Report
      2021 Research-status Report
  • [Remarks] 臨床検体中のコロナウイルスタンパク質を蛍光抗体で迅速定量することに成功

    • URL

      https://www.titech.ac.jp/news/2022/065515

    • Related Report
      2022 Research-status Report
  • [Remarks] Quenchbodies Pave the Way to COVID-19 Diagnostics

    • URL

      https://www.titech.ac.jp/english/news/2022/065498

    • Related Report
      2022 Research-status Report

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Published: 2021-04-28   Modified: 2025-01-30  

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