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Development of efficient methods for preparation of high-affinity antibody mutants suitable for sensitive detection of small biomarker molecules

Research Project

Project/Area Number 26860378
Research Category

Grant-in-Aid for Young Scientists (B)

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

Principal Investigator

Oyama Hiroyuki  神戸薬科大学, 薬学部, 助教 (80572966)

Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords抗体工学 / 臨床化学 / タンパク質工学 / 進化分子工学 / 免疫化学
Outline of Final Research Achievements

Antibodies with high affinity and specificity work as a powerful tool for in vitro and in vivo clinical diagnosis. These antibodies have been prepared by the hybridoma method using immune spleen cells, but often lack sufficient binding abilities. Recent antibody engineering technology could improve the native antibodies, but considerable labor and period are required to select rare improved binders from a vast library of various mutated antibodies. To overcome this problem, this study aimed to develop innovative methods for isolating the improved species based on the flow cytometry principle. We prepared a key reagent for these systems, a monoclonal anti-phage antibody, and converted it to relevant single-chain Fv fragment (scFv). Moreover, this scFv was linked with a luminescent protein, and utility of this fusion protein was investigated.

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • Research Products

    (13 results)

All 2016 2015 2014

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Presentation (11 results) (of which Int'l Joint Research: 1 results,  Invited: 1 results) Book (1 results)

  • [Journal Article] Gaussia Luciferase as a Genetic Fusion Partner with Antibody Fragments for Sensitive Immunoassay Monitoring of Clinical Biomarkers2015

    • Author(s)
      Oyama Hiroyuki; Morita Izumi; Kiguchi Yuki; Miyake Sayaka; Moriuchi Ayaka; Akisada Tatsuki; Kobayashi Norihiro; Niwa Toshifumi
    • Journal Title

      Analytical chemistry

      Volume: 87(24) Issue: 24 Pages: 12387-12395

    • DOI

      10.1021/acs.analchem.5b04015

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Presentation] 抗コルチゾールscFvの親和性成熟における部位特異的点変異の効果2016

    • Author(s)
      大山浩之, 德重仁美, 知念絵美里, 森下知美, 森田いずみ, 小林典裕
    • Organizer
      日本薬学会第136年会
    • Place of Presentation
      横浜
    • Year and Date
      2016-03-28
    • Related Report
      2015 Annual Research Report
  • [Presentation] 試験管内親和性成熟による実用抗体の創製: 低分子バイオマーカーを例に2015

    • Author(s)
      大山浩之
    • Organizer
      生物化学的測定研究会第20回学術シンポジウム
    • Place of Presentation
      神戸
    • Year and Date
      2015-11-13
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 高親和力抗エストラジオール変異scFvの創製とその親和性成熟機構の解析2015

    • Author(s)
      大山浩之, 江浪友理, 山本知佳, 田川達矢, 小林典裕
    • Organizer
      第55回日本臨床化学会年次学術集会
    • Place of Presentation
      吹田
    • Year and Date
      2015-10-31
    • Related Report
      2015 Annual Research Report
  • [Presentation] 改良型抗エストラジオールscFvの親和性成熟機構の解析2015

    • Author(s)
      大山浩之, 江浪友理, 田川達矢, 山本知佳, 森田いずみ, 小林典裕
    • Organizer
      日本分析化学会64年会
    • Place of Presentation
      博多
    • Year and Date
      2015-09-11
    • Related Report
      2015 Annual Research Report
  • [Presentation] “Antibody breeding” for more sensitive immunoassays 1: Three-step affinity maturation generated an improved scFv suitable for serum estradiol-17 beta ELISA2015

    • Author(s)
      Hiroyuki Oyama, Norihiro Kobayashi
    • Organizer
      European Congress of Clinical Chemistry and Laboratory Medicine
    • Place of Presentation
      Paris, France
    • Year and Date
      2015-06-22
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 抗エストラジオールscFv試験管内親和性成熟機構の解析2015

    • Author(s)
      大山浩之, 江浪友理, 木口裕貴, 森田いずみ, 小林典裕
    • Organizer
      日本薬学会第135年会
    • Place of Presentation
      神戸市
    • Year and Date
      2015-03-27
    • Related Report
      2014 Research-status Report
  • [Presentation] 抗コチニンscFvの親和性成熟における部位特異的点変異の効果2015

    • Author(s)
      森田いずみ, 西村沙貴, 蓑田早耶香, 木口裕貴, 大山浩之, 小林典裕
    • Organizer
      日本薬学会第135年会
    • Place of Presentation
      神戸市
    • Year and Date
      2015-03-27
    • Related Report
      2014 Research-status Report
  • [Presentation] アミロイドーシスの病態解明を目指した抗ApoA-Iモノクローナル抗体の作製2015

    • Author(s)
      堀江有紀, 土屋沙織, 大山浩之, 小林典裕, 斎藤博幸
    • Organizer
      日本薬学会第135年会
    • Place of Presentation
      神戸市
    • Year and Date
      2015-03-27
    • Related Report
      2014 Research-status Report
  • [Presentation] 大麻成分に対するモノクローナル抗体の調製とキャラクタリゼーション2015

    • Author(s)
      伊藤綾, 安尾まゆみ, 竜田都加, 森田いずみ, 大山浩之, 小林典裕
    • Organizer
      日本薬学会第135年会
    • Place of Presentation
      神戸市
    • Year and Date
      2015-03-27
    • Related Report
      2014 Research-status Report
  • [Presentation] 高感度ヒト血清中コルチゾールELISAの開発を目的とするscFv-ルシフェラーゼ融合タンパク質の創製2014

    • Author(s)
      大山浩之, 三宅沙也香, 森田いずみ, 丹羽俊文, 小林典裕
    • Organizer
      日本分析化学会第63年会
    • Place of Presentation
      東広島市
    • Year and Date
      2014-09-19
    • Related Report
      2014 Research-status Report
  • [Presentation] 改良型変異scFvフラグメントを用いるヒト尿中コチニンのモノクローナルELISA2014

    • Author(s)
      森田いずみ, 平井杏奈, 西村沙貴, 大山浩之, 太田光煕, 小林典裕
    • Organizer
      日本分析化学会第63年会
    • Place of Presentation
      東広島市
    • Year and Date
      2014-09-19
    • Related Report
      2014 Research-status Report
  • [Book] 免疫測定法 基礎から先端まで2014

    • Author(s)
      生物化学的測定研究会(編)
    • Total Pages
      336
    • Publisher
      講談社
    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2017-05-10  

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