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Generation and clinical application of low-molecular-weight functional domain proteins.

Research Project

Project/Area Number 25670280
Research Category

Grant-in-Aid for Challenging Exploratory Research

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

Principal Investigator

KOBAYASHI NORIHIRO  神戸薬科大学, 薬学部, 教授 (90205477)

Co-Investigator(Kenkyū-buntansha) OYAMA Hiroyuki  神戸薬科大学, 薬学部, 助教 (80572966)
MORITA Izumi  神戸薬科大学, 薬学部, 助手 (20299085)
Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords進化分子工学
Outline of Final Research Achievements

Although antibodies are essential for trace characterization of diagnostic biomarkers, their high molecular weight and low expression levels in E.coli make it difficult to improving their functions by genetic engineering. To overcome these limitations, we planned generation of low-molecular-weight functional domain proteins. First, we made a molecular library, each member of which has the VH domain of an antibody as a fundamental structure and a randomized complementarity-determining region 3 loop, but any practical species could not be isolated. Then, another library based on the streptavidin monomer was produced. Three loop structures, which were supposed to contact with steroids by in silico modeling, were selected as the target of randomization. This library provided the species that were evaluated to had gained the ability to recognize and bind estradiol or cortisol.

Report

(3 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • Research Products

    (17 results)

All 2015 2014 2013 Other

All Journal Article (3 results) (of which Peer Reviewed: 3 results,  Acknowledgement Compliant: 1 results) Presentation (12 results) Book (1 results) Remarks (1 results)

  • [Journal Article] Direct detection of ABCA1-dependent HDL formation based on lipidation-induced hydrophobicity change in apoA-I.2014

    • Author(s)
      Risa Omura, Kohjiro Nagao, Norihiro Kobayashi, Kazumitsu Ueda, and Hiroyuki Saito
    • Journal Title

      J Lipid Res.

      Volume: 55 Issue: 11 Pages: 2423-2431

    • DOI

      10.1194/jlr.d049445

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Anti-idiotype scFv-enzyme fusion proteins: A clonable analyte-mimicking probe for standardized immunoassays targeting small biomarkers.2013

    • Author(s)
      Oyama H., Tanaka E., Kawanaka T., Morita I., Niwa T., Kobayashi N.
    • Journal Title

      Analytical Chemistry

      Volume: 85 Issue: 23 Pages: 11553-11559

    • DOI

      10.1021/ac402868f

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] “Breeding” diagnostic antibodies for higher assay performance: a 250-fold affinity-matured antibody mutant targeting a small biomarker.2013

    • Author(s)
      Oyama H., Yamaguchi S., Nakata S., Niwa T., Kobayashi N.
    • Journal Title

      Analytical Chemistry

      Volume: 85 Issue: 10 Pages: 4930-4937

    • DOI

      10.1021/ac3037602

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [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,江波 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 Annual Research 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 Annual Research 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 Annual Research 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 Annual Research Report
  • [Presentation] バイオレイヤー干渉法を用いた人工 HDL 粒子と抗 apoA-I 抗体との相互作用評価2015

    • Author(s)
      木村 仁,武知 佑樹,小林 典裕,齋藤 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 Annual Research 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 Annual Research 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 光熙,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 Annual Research Report
  • [Presentation] 試験管内親和性成熟scFvを用いるヒト尿中コチニンのELISA2013

    • Author(s)
      大山 浩之,森田 いずみ,太田 光熙,小林 典裕
    • Organizer
      生物化学的測定研究会 第18回学術集会・総会
    • Place of Presentation
      京都
    • Related Report
      2013 Research-status Report
  • [Presentation] 高親和力scFv融合タンパク質を用いるコルチゾールの高感度ELISA2013

    • Author(s)
      大山 浩之,森田 いずみ,太田 光熙,小林 典裕
    • Organizer
      第53回日本臨床化学会年次学術集会
    • Place of Presentation
      徳島
    • Related Report
      2013 Research-status Report
  • [Presentation] Anti-Idiotype scFv-Enzyme Fusion Proteins Work as a Clonable Analyte-Mimicking Probe That Enables Standardization of Hapten Immunoassays2013

    • Author(s)
      Kobayashi N., Oyama H., Morita I., Niwa T.
    • Organizer
      Fifth Annual PEGS Europe : Protein & Antibody Engineering Summit
    • Place of Presentation
      Lisbon
    • Related Report
      2013 Research-status Report
  • [Presentation] "Breeding" Diagnostic Antibodies: A 250-Fold Affinity-Matured scFv Mutant for Measuring Human Serum Estradiol-17β2013

    • Author(s)
      Oyama H., Niwa T., Kobayashi N.
    • Organizer
      Fifth Annual PEGS Europe : Protein & Antibody Engineering Summit
    • Place of Presentation
      Lisbon
    • Related Report
      2013 Research-status Report
  • [Presentation] "Breeding" Diagnostic Antibodies: An Affinity-Matured scFv for Urinary Cotinine ELISA to Monitor Tobacco Smoke Exposure2013

    • Author(s)
      Morita I., Oyama H., Banzono E., Ohta M., Kobayashi N.
    • Organizer
      Fifth Annual PEGS Europe : Protein & Antibody Engineering Summit
    • Place of Presentation
      Lisbon
    • Related Report
      2013 Research-status Report
  • [Book] 免疫測定法2014

    • Author(s)
      生物化学的測定研究会
    • Total Pages
      325
    • Publisher
      講談社
    • Related Report
      2014 Annual Research Report
  • [Remarks] 神戸薬科大学 生命分析化学研究室HP

    • URL

      http://www.kobepharma-u.ac.jp/analchem/

    • Related Report
      2014 Annual Research Report

URL: 

Published: 2014-07-25   Modified: 2019-07-29  

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