• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Development of immunoassay for fish vitellogenin using universal antibody

Research Project

Project/Area Number 11556038
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section展開研究
Research Field Fisheries chemistry
Research InstitutionHokkaido University

Principal Investigator

HARA Akihiko  Hokkaido Univ., Grad.School of Fisheries Sciences, Prof., 水産科学研究科, 教授 (40091483)

Co-Investigator(Kenkyū-buntansha) FUJINO Hiroyoshi  Katayama Chem.Indus.Co.Ltd, ASection Chief, 研究開発部, 課長(研究職)
IGUCHI Kazuaki  Univ., Shizuoka, School of Pharmaceutical Sci., Lec., 薬学部, 講師 (20176077)
ADACHI Shinji  Hokkaido Univ., Grad.School of Fisheries Sciences, Asso.Prof., 水産科学研究科, 助教授 (40231930)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 2000: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1999: ¥4,200,000 (Direct Cost: ¥4,200,000)
Keywordsvitellogenin / lipovitellin / β'-component / salmond / cyprinid / enzyme immunoassay / chemiluminescent immunoassay / 免疫測定法 / EIA / CLIA / β'-コンポーネント / モノクロナール抗体 / サケマス / コイ / 酵素免疫測定法
Research Abstract

In order to establish universal immunoassay for fish vitellogenins as a biomarker of environmental estrogens, we developed sensitive enzyme immunoassay(EIA)and chemiluminescent immunoassay(CLIA)using monoclonal and/or polyclonal antibodies again st purified egg yolk proteins from salmon and carp.
1. Generation of antibodies against egg yolk proteins : Salmon egg yolk proteins are composed of three components : lipovitellin, phosvitin and β'-component(beta). Specific antibodies in rabbits and mice against purified masu salmon lipovitellin and beta, as well as against carp lipovitellin were generated. Beta in carp was not detected in this study.
2. Amino acid sequence of masu salmon beta : N-terminal amino acid sequence of masu salmon beta was analyzed and 35 residues were determined. The amino acid sequence was in good agreement with that of rainbow trout previously reported.
3. Development of vitellogenin immunoassays : The CLIA for carp vitellogenin was completed within foru hours and measured vitellogenin from 2 to 1000 ng/ml. Cross-reactivity was found in cyprinids, but not in salmonids. Also EIA and CLIA for salmon vitellogenin were developed. As for the EIA, sandwich method using polyclonal and monoclonal antibodies against masu salmon lipovitellin and β'-component, respectively, was used. The EIA measures vitellogenin as low as 0.3 ng/ml. The CLIA developed in this study is called one-step CLIA using two types of antibodies. The CLIA was completed within 6 to 9 hours and covered a range of 122pg/ml to 1000 ng/ml. Both EIA and CLIA for salmon vitellogenin cross-reacted with vitellogenins from other salmonids and therefore can be used as universal immunoassays for salmon vitellogenins.
4. Development of commercial vitellogenin assay kit : For the primary screening test prior to analysis by EIA and CLIA, single radial immunodiffusion was developed. This plate is able to measure vitellogenin in a range of 25 to 1000 mg/ml and is ready to be on the market.

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] 原彰彦: "水環境における汚染影響評価のバイオマーカーとしてのビテロジェニン"日本環境毒性学会誌. 2. 35-42 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 原彰彦: "内分泌撹乱物質の生態影響-魚類への影響"廃棄物学会誌. 10. 278-287 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Hiramatsu: "Simple and sensitive detection of vitellogenin receptor (s) in Sakhalin taimen (Hucho perryi)."Bull.Fish.Sci.Hokkaido Univ.. 52. 5-9 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Fukada: "Changes of serum growth hormone and vitellogenin levels during vitellogenesis in female masu salmon (Oncorhynchus masou)."Fish.Sci.. 66. 789-791 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 原彰彦: "魚類の卵形成と雌特異蛋白質ビテロジェニン-環境ホルモンのバイオマーカー"化学と生物. 39. 29-36 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hara, A.: "Vitellogenin in male as a biomarker for estrogenic contamination of the aquatic environment"Jpn.J.Environ.Toxicol.. 2(1). 35-42 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hara, A.: "Effects on wildlife exposed to endocrine disrupters with special reference to fishes"Waste Manage.Res.. 10(4). 278-287 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hiramatsu, N.et al.: "Simple and sensitive detection of vitellogenin receptor(s)in Sakhalin taimen(Hucho perryi)"Bull.Fish.Sci.Hokkaido Univ.. 52. 5-9 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Fukada, H.et al.: "Changes of serum growth hormone and vitellogenin levels during vitellogenesis in female masu salmon(Oncorhynchus masou)"Fish.Sci.. 66. 789-791 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hara, A.: "Vitellogenesis and female-specific serum proteins "vitellogenin" in fish-vitellogenin as a biomarker for estrogenic contamination of the aquatic environment"Kagaku To Seibutsu. 39(1). 29-36 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Hiramatsu: "Simple and sensitive detection of vitellogenin receptor (s) in Sakhalin taimen (Hucho perryi) ."Bull.Fish.Sci.Hokkaido Univ.. 52・1. 5-9 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] H.Fukada: "Changes of serum growth hormone and vitellogenin levels during vitellogenesis in female masu salmon (Oncorhynchus masou) ."Fish.Sci.. 66・4. 789-791 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 原彰彦: "魚類の卵形成と雌特異蛋白質ビテロジェニン-環境ホルテンのバイオマーカー"化学と生物. 39・1. 29-36 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 原 彰彦: "水環境における汚染影響評価のバイオマーカーとしてのビテロジェニン"日本環境毒性学会誌. 2・1. 35-42 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 原 彰彦: "魚類ビテロジェニン研究と環境ホルモン"医学のあゆみ. 190・7〜8. 745-746 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 原 彰彦: "内分泌撹乱物質の生態影響-魚類への影響"廃棄物学会誌. 10・4. 278-287 (1999)

    • Related Report
      1999 Annual Research Report

URL: 

Published: 1999-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi