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Development of new immunoassay systems for on-site analysis targeting stimulant amines

Research Project

Project/Area Number 16K08953
Research Category

Grant-in-Aid for Scientific Research (C)

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

Principal Investigator

MORITA IZUMI  神戸薬科大学, 薬学部, 助教 (20299085)

Co-Investigator(Kenkyū-buntansha) 小林 典裕  神戸薬科大学, 薬学部, 教授 (90205477)
大山 浩之  神戸薬科大学, 薬学部, 講師 (80572966)
上田 宏  東京工業大学, 科学技術創成研究院, 教授 (60232758)
Research Collaborator YOSHIDA TAKEMI  
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: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords抗体 / 覚せいアミン / メタンフェタミン / 第Ⅱ相代謝物 / モノクローナル抗体 / ELISA / アンフェタミン / 誘導体化 / 臨床化学
Outline of Final Research Achievements

For law enforcement against illicit use of stimulant amines, i.e., amphetamine (AP) and methamphetamine (MAP), rapid and feasible on-site assays are required. Although immunochemical approach with specific antibodies is promising, practical antibodies for these stimulant amines are not available now, and difficult to generate because of their small molecular weight. In this study, we generated novel monoclonal antibodies that enable detection of AP, MAP, and the urinary MAP metabolite, p-hydroxy-MAP sulfate (MAPOH-S), by immunizing mice with newly designed immunogenic conjugates. For detecting AP and MAP, antibodies were elicited against silyl derivatives of AP and MAP that show more favorable immunogenicity due to enlarged molecular mass, whereas antibodies recognizing MAPOH-S itself (without modification) were obtained. We expect that these antibodies may allow us to develop practical on-site analytical systems, e.g., immunochromatographies, targeting AP, MAP, and MAPOH-S.

Academic Significance and Societal Importance of the Research Achievements

覚せいアミン類の分析には、現在、可搬性に乏しい大型機器を用いる機器分離分析が多用されているが、複雑な試料の前処理が必須で多大の時間と労力を要する。簡便・迅速で高感度な現場分析を可能にすることは、覚せい剤乱用に係る社会的問題の大きさを鑑みれば極めて意義が大きい。今回、特に分子量が小さく高親和力抗体の調製が難しい覚せいアミン類について、誘導体化も含めた免疫原の工夫により、実用的な親和力を持つモノクローナル抗体を樹立することができた。本研究の基盤となった戦略と技術は、他の規制薬物、環境汚染物質、臨床バイオマーカーなど、低分子量物質のオンサイト免疫測定法の確立にも広く適用が可能と期待される。

Report

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

    (21 results)

All 2019 2018 2017 2016

All Journal Article (6 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Open Access: 3 results) Presentation (15 results) (of which Int'l Joint Research: 2 results,  Invited: 1 results)

  • [Journal Article] Antibodies and Engineered Antibody Fragments against M13 Filamentous Phage to Facilitate Phage-Display-Based Molecular Breeding2018

    • Author(s)
      Y. Kiguchi, H. Oyama, I. Morita, E. Katayama, M. Fujita, M. Narasaki, A. Yokoyama, N. Kobayashi
    • Journal Title

      Biological and Pharmaceutical Bulletin

      Volume: 41 Issue: 7 Pages: 1062-1070

    • DOI

      10.1248/bpb.b18-00162

    • NAID

      130007387201

    • ISSN
      0918-6158, 1347-5215
    • Year and Date
      2018-07-01
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Enantioselective Monoclonal Antibodies for Detecting Ketamine to Crack Down on Illicit Use2018

    • Author(s)
      Morita, I. Oyama, H. Kanda, Y. Yasuo, M. Ito, A. Toyota, M. Hayashi, Y. Yokoyama, T. Kobayashi, N.
    • Journal Title

      Biological and Pharmaceutical Bulletin

      Volume: 41 Issue: 1 Pages: 123-131

    • DOI

      10.1248/bpb.b17-00762

    • NAID

      130006301299

    • ISSN
      0918-6158, 1347-5215
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Antibody Fragments for On-Site Testing of Cannabinoids Generated <i>via in Vitro</i> Affinity Maturation2017

    • Author(s)
      Morita Izumi; Oyama Hiroyuki; Yasuo Mayumi; Matsuda Kazuhisa; Katagi Kengo; Ito Aya; Tatsuda Hiroka; Tanaka Hiroyuki; Morimoto Satoshi; Kobayashi Norihiro
    • Journal Title

      Biological and Pharmaceutical Bulletin

      Volume: 40 Issue: 2 Pages: 174-181

    • DOI

      10.1248/bpb.b16-00669

    • NAID

      130005308428

    • ISSN
      0918-6158, 1347-5215
    • Related Report
      2017 Research-status Report 2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] A Single-Step “Breeding” Generated a Diagnostic Anti-cortisol Antibody Fragment with Over 30-Fold Enhanced Affinity2017

    • Author(s)
      H. Oyama, I. Morita, Y. Kiguchi, T. Morishita, S. Fukushima, Y. Nishimori, T. Niwa, N. Kobayashi
    • Journal Title

      Biological and Pharmaceutical Bulletin

      Volume: 40 Issue: 12 Pages: 2191-2198

    • DOI

      10.1248/bpb.b17-00633

    • NAID

      130006235440

    • ISSN
      0918-6158, 1347-5215
    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] One-Shot In Vitro Evolution Generated an Antibody Fragment for Testing Urinary Cotinine with More Than 40-Fold Enhanced Affinity2017

    • Author(s)
      Oyama, Hiroyuki; Morita, Izumi; Kiguchi, Yuki; Banzono, Erika; Ishii, Kasumi; Kubo, Satoshi; Watanabe, Yoshiro; Hirai, Anna; Kaede, Chiaki; Ohta, Mitsuhiro; Kobayashi Norihiro
    • Journal Title

      Analytical Chemistry

      Volume: 89(1) Issue: 1 Pages: 988-995

    • DOI

      10.1021/acs.analchem.6b04332

    • Related Report
      2017 Research-status Report 2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] Immunochemical Approach for Monitoring of Structural Transition of ApoA-I upon HDL Formation Using Novel Monoclonal Antibodies2017

    • Author(s)
      H. Kimura, S. Mikawa, C. Mizuguchi, Y. Horie, I. Morita, H. Oyama, T. Ohgita, K. Nishitsuji, A. Takeuchi, S. Lund-Katz, K. Akaji, N. Kobayashi, H. Saito
    • Journal Title

      Sci. Rep.

      Volume: 7 Issue: 1 Pages: 2988-2988

    • DOI

      10.1038/s41598-017-03208-8

    • NAID

      120006534990

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] メタンフェタミン第Ⅱ相代謝物に対する特異モノクローナル抗体産生の試み2019

    • Author(s)
      崎尾 奈美, 森田 いずみ, 柏原 圭佑, 横田 朝香, 大山 浩之, 吉田 武美, 上田 宏, 小林 典裕
    • Organizer
      日本薬学会第139年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 幻覚性キノコ成分シロシンのオンサイト分析を目的とする新規モノクローナル抗体の作製2018

    • Author(s)
      森田 いずみ、大山 浩之、田中 理恵、花尻(木倉) 瑠理、小林 典裕
    • Organizer
      日本法中毒学会第37年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 抗Δ9-テトラヒドロカンナビノールscFvの部位特異的変異導入による試験管内親和性成熟の試み2018

    • Author(s)
      大山 浩之、肥越 ひかる、小林 奈津子、田口 歩維、比嘉 ひかり、森田 いずみ、小林 典裕
    • Organizer
      日本法中毒学会第37年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 新世代の高性能イムノアッセイを目指す「抗体育種」2018

    • Author(s)
      小林 典裕、大山 浩之、森田 いずみ、木口 裕貴
    • Organizer
      日本分析化学会第67年会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] In vitro affinity maturation of a single-chain Fv fragment for on-site testing of cannabinoids2017

    • Author(s)
      Izumi Morita, Hiroyuki Oyama, Hiroyuki Tanaka, Satoshi Morimoto, and Norihiro Kobayashi
    • Organizer
      European Congress of Clinical Chemistry and Laboratory Medicine
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Utility of Gaussia luciferase as a fusion partner with scFvs for bioluminescent immunoassays testing clinical biomarkers2017

    • Author(s)
      Hiroyuki Oyama, Izumi Morita, Toshifumi Niwa, and Norihiro Kobayashi
    • Organizer
      European Congress of Clinical Chemistry and Laboratory Medicine
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] 新規モノクローナル抗ケタミン抗体の作製とその諸性質2017

    • Author(s)
      森田いずみ, 大山浩之, 神田結, 安尾まゆみ, 伊藤綾, 小林典裕
    • Organizer
      日本法中毒学会第36年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 抗Δ9-テトラヒドロカンナビノール一本鎖Fvフラグメントの作製と試験管内親和性成熟2017

    • Author(s)
      大山浩之, 森田いずみ, 松田和久, 片木謙吾, 小林典裕, 田中宏幸, 森元聡
    • Organizer
      日本法中毒学会第36年会
    • Related Report
      2017 Research-status Report
  • [Presentation] アッセイ感度の向上を目指す「抗体育種」:抗コチニン抗体を例として2017

    • Author(s)
      小林 典裕、大山 浩之、森田 いずみ、木口 裕貴
    • Organizer
      日本法中毒学会第36年会
    • Related Report
      2017 Research-status Report
  • [Presentation] オンサイト免疫測定法を目的としたモノクローナル抗ケタミン抗体の新規調製2017

    • Author(s)
      森田いずみ, 神田結, 安尾まゆみ, 伊藤綾, 大山浩之, 小林典裕
    • Organizer
      日本分析化学会66年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 高親和力を保持した抗エストラジオールscFv最少変異体調製の試み2017

    • Author(s)
      7.大山浩之, 木口裕貴, 井上由香莉, 高嶺百合子, 藤原若菜, 森田いずみ, 小林典裕
    • Organizer
      日本分析化学会66年会
    • Related Report
      2017 Research-status Report
  • [Presentation] ApoA-I構造特異抗体の開発に向けた抗体評価系の確立2017

    • Author(s)
      三河志穂、木村仁、森田いずみ、大山浩之、小林典裕、斎藤博幸
    • Organizer
      日本薬学会第137年会
    • Place of Presentation
      仙台
    • Related Report
      2016 Research-status Report
  • [Presentation] Effect of Heparin on Amyloid Fibril Formation of ApoA-I Fragment Peptides2016

    • Author(s)
      Shiho Mikawa, Chiharu Mizuguchi, Izumi Morita, Hioryuki Oyama, Teruhiko Baba, Akira Shigenaga, Toshinori Shimanouchi, Norihiro Kobayashi, Akira Otaka, Kenichi Akaji, Hiroyuki Saito
    • Organizer
      第53回ペプチド討論会
    • Place of Presentation
      京都
    • Year and Date
      2016-10-26
    • Related Report
      2016 Research-status Report
  • [Presentation] オンサイト免疫測定法を目的としたTeoc化覚せいアミンに対するモノクローナル抗体の新規調製2016

    • Author(s)
      森田いずみ、黒田裕美、伊藤綾、池田夏美、小山千尋、大山浩之、小林典裕
    • Organizer
      日本分析化学会第65年会
    • Place of Presentation
      札幌
    • Related Report
      2016 Research-status Report
  • [Presentation] 大麻成分の検出を目的とする抗Δ9-テトラヒドロカンナビノールscFvの試験管内親和性成熟2016

    • Author(s)
      大山浩之、森田いずみ、松田和久、伊藤綾、小林典裕
    • Organizer
      日本分析化学会第65年会
    • Place of Presentation
      札幌
    • Related Report
      2016 Research-status Report

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

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