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Development of a mass spectrometry-based quantitative assay for hepatitis virus detection

Research Project

Project/Area Number 16K08937
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Laboratory medicine
Research InstitutionEhime University

Principal Investigator

Takemori Nobuaki  愛媛大学, 学術支援センター, 講師 (40533047)

Co-Investigator(Kenkyū-buntansha) 鈴木 哲朗  浜松医科大学, 医学部, 教授 (00250184)
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords質量分析 / 選択反応モニタリング / バイオマーカー / 肝炎ウイルス / ウイルス
Outline of Final Research Achievements

Serological diagnosis of hepatitis B virus is commonly performed with immunoassays against the HBV surface antigen (HBsAg). However, an alternative approach for HBsAg detection is currently required because of the emergence of immune-escape mutations, which cause false negative results in diagnostic tests. In this study, we developed an antibody-free strategy for HBsAg quantitation using targeted proteomics. By profiling tryptic HBsAg digests, we identified signature peptides that enabled the establishment of a reliable targeted assay for HBsAg quantitation, using selected reaction monitoring mass spectrometry. The established assay detected HBsAg at the sub-picogram level in a 100-nanoliter serum sample, and parallel monitoring of amino acid variations in signature peptides facilitated the determination of HBsAg genotypes. Our results demonstrate the feasibility of mass spectrometry-based diagnosis of HBV infection as an alternative method to conventional antibody-based diagnosis.

Academic Significance and Societal Importance of the Research Achievements

質量分析をベースとした検査法を新たに開発することで、従来のイムノアッセイ法で問題とされてきたエスケープ変異体による感染リスクの低減が可能となり、肝炎ウイルス感染の拡大防止に寄与できる。将来的には、新規ウイルスや、イムノアッセイの確立されていないウイルスに対する質量分析ベースの血液診断法を確立することにより感染症一般への対策に貢献する。

Report

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

    (11 results)

All 2019 2018 2017 2016

All Journal Article (4 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 4 results,  Open Access: 3 results) Presentation (6 results) (of which Int'l Joint Research: 6 results,  Invited: 1 results) Patent(Industrial Property Rights) (1 results) (of which Overseas: 1 results)

  • [Journal Article] Quantitative assay of targeted proteome in tomato trichome glandular cells using a large-scale selected reaction monitoring strategy2019

    • Author(s)
      Takemori Ayako、Nakashima Taiken、Oomura Hisashi、Tanaka Yuki、Nakata Keisuke、Nonami Hiroshi、Takemori Nobuaki
    • Journal Title

      Plant Methods

      Volume: 15 Issue: 1 Pages: 40-40

    • DOI

      10.1186/s13007-019-0427-7

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A Multiplexed Co-synthesis System for Stable Isotope-labeled Peptide Standards Using Wheat Germ Cell-free Synthesis and Its Application to Quantitative Proteomics2018

    • Author(s)
      武森 信曉
    • Journal Title

      Proteome Letters

      Volume: 3 Issue: 1 Pages: 23-30

    • DOI

      10.14889/jpros.3.1_23

    • NAID

      130007402397

    • ISSN
      2432-2776
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] MEERCAT: Multiplexed Efficient Cell Free Expression of Recombinant QconCATs For Large Scale Absolute Proteome Quantification2017

    • Author(s)
      Takemori Nobuaki、Takemori Ayako、Tanaka Yuki、Endo Yaeta、Hurst Jane L.、Gomez-Baena Guadalupe、Harman Victoria M.、Beynon Robert J.
    • Journal Title

      Molecular & Cellular Proteomics

      Volume: 16 Issue: 12 Pages: 2169-2183

    • DOI

      10.1074/mcp.ra117.000284

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Top-down/Bottom-up Mass Spectrometry Workflow Using Dissolvable Polyacrylamide Gels2017

    • Author(s)
      Takemori Nobuaki、Takemori Ayako、Wongkongkathep Piriya、Nshanian Michael、Loo Rachel R. Ogorzalek、Lermyte Frederik、Loo Joseph A.
    • Journal Title

      Analytical Chemistry

      Volume: 89 Issue: 16 Pages: 8244-8250

    • DOI

      10.1021/acs.analchem.7b00357

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] A multiplexed co-synthesis system for stable isotope-labeled peptide standards using wheat germ cell-free synthesis and its application to quantitative proteomics (JPrOS Award Lecture)2018

    • Author(s)
      Nobuaki Takemori
    • Organizer
      Mass Spectrometry and Proteomics 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] High-throughput biosynthesis of stable isotope-labeled internal standards for global proteome quantification,2018

    • Author(s)
      Nobuaki Takemori
    • Organizer
      Mass Spectrometry and Proteomics 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] High-speed recovery workflow for electrophoretically separated proteins in polyacrylamide gels and its applications to mass spectrometry2018

    • Author(s)
      Ayako Takemori, Victoria M Harman, Philip Brownridge, Joseph A. Loo, Rachel R. Ogorzalek Loo, Robert J Beynon, and Nobuaki Takemori
    • Organizer
      HUPO 17th Annual World Congress in Orlando
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Proteomics workflow using dissolvable polyacrylamide gels2017

    • Author(s)
      Takemori Nobuaki、Takemori Ayako、Wongkongkathep Piriya、Nshanian Michael、Loo Rachel R. Ogorzalek、Lermyte Frederik、Wu Si、Loo Joseph A.
    • Organizer
      Twelfth International Symposium on Mass Spectrometry in the Health & Life Science: Molecular & Cellular Proteomics
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Multiple QconCAT biosynthesis in a cell-free protein synthesis system2017

    • Author(s)
      Takemori Nobuaki、Takemori Ayako、Tanaka Yuki、Endo Yaeta、Hurst Jane L.、Gomez-Baena Guadalupe、Harman Victoria M.、Beynon Robert J.
    • Organizer
      Twelfth International Symposium on Mass Spectrometry in the Health & Life Science: Molecular & Cellular Proteomics
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Top-down Mass Spectrometry using BAC-PAGE2017

    • Author(s)
      Takemori Nobuaki、Takemori Ayako、Wongkongkathep Piriya、Nshanian Michael、Loo Rachel R. Ogorzalek、Lermyte Frederik、Wu Si、Loo Joseph A.
    • Organizer
      7th Asia-Oceania Mass Spectrometry Conference
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Patent(Industrial Property Rights)] 質量分析を用いたB型肝炎ウイルスの測定方法2016

    • Inventor(s)
      武森信暁、武森文子、鈴木哲朗
    • Industrial Property Rights Holder
      愛媛大学、浜松医科大学
    • Industrial Property Rights Type
      特許
    • Filing Date
      2016-07-22
    • Related Report
      2016 Research-status Report
    • Overseas

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

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