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Development of high sensitive fluoroimmunoassay for synthetic adrenal agents and its clinical application

Research Project

Project/Area Number 63571116
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Laboratory medicine
Research InstitutionKobe women's College of Pharmacy

Principal Investigator

WATANABE Fukuko  Kobe Women's College of Pharmacy, Faculty of Pharmacy, Professor, 薬学部, 教授 (10068322)

Co-Investigator(Kenkyū-buntansha) NISHIGUCHI Yoshino  Kobe Women's College of Pharmacy, Faculty of Pharmacy, Assistant, 薬学部, 助手 (60208171)
KIGUCHI Toshiko  Kobe Women's College of Pharmacy, Faculty of Pharmacy, Lecturer, 薬学部, 講師 (70068344)
KOBAYASHI Yoshiharu  Kobe Women's College of Pharmacy, Faculty of Pharmacy, Lecturer, 薬学部, 講師 (20133647)
NAITOH Takeaki  Kobe Women's College of Pharmacy, Faculty of Pharmacy, Professor, 薬学部, 教授 (00068339)
NINOMIYA Ichiya  Kobe Women's College of Pharmacy, Faculty of Pharmacy, Professor, 薬学部, 教授 (00068321)
多河 典子  神戸女子薬科大学, 薬学部, 助手 (00206905)
Project Period (FY) 1988 – 1989
Project Status Completed (Fiscal Year 1989)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1989: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1988: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsFluorescence polarization immunoassay / Dexamethasone / Prednisolone / FITC-labled steroid / 蛍光免疫定量法 / プレドニゾロン、
Research Abstract

In this study, the new approach to establishment of the high sensitive fluorescence polarization immunoassay system of the synthetic adrenal agents such as dexamethasone and prednisolone are achieved. Argon-ion laser as a light source of a model IBF-129 fluorescence polarization fluorimeter, sample changer, and micro flow-cell were used in this semi-automatic system.
These immunoassays require the preparation of effective antibody and FITC-labeled antigen, thus necessitating the synthesis of appropriate of steroids, carying substituent capable of binding with protein or FITC. For this purpose, we were synthesized 4-(carboxymethylthio)-dexamethasone and prednisolone-21-hemisuccinate. These steroid derivatives werd conjugated with bovine serum albumin (BSA) by mixed anhydride reaction, and the antibody to the conjugate were raised in rabbits. The purified IgG fraction of steroid antiserum was prepared by applying to DEAE-Sephacel column. Cross- reactivity of cortisol to anti-prednisolone-21-hemisuccinate-BSA antiserum was 9.6%, specificity of this antibody was quite similar to those of the previous reports of some investigators.
FITC-labeled antigen are prepared by conjugation the derivative steroids with the fluorescein isothiocyanate.
Procedure: Serum sample was mixed with FITC-labeled steroid and polarization value (A) was measured. Then anti-steroid antiserum IgG fraction was added. The mixture was incubated at constant temperature of 20-25 ゚C for 60 min, and the polarization value (B) was measured again. A correction was made for minimizing interference due to serum by subtracting the polarization value(A) from the value (B).

Report

(3 results)
  • 1989 Annual Research Report   Final Research Report Summary
  • 1988 Annual Research Report

Research Products

(10 results)

All Other

All Publications (10 results)

  • [Publications] Yoshino NISHIGUGHI: "Synthesis of 4-Carboxymethylthio-Substituted Steroids for Enzyme Immunoassay." Chem.Pharm.Bull.(Tokyo).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 小林吉晴: "レ-ザ-励起による蛍光偏光免疫定量法の高感度化" 臨床化学.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Fukuko WATANABE: "NONISOTOPIC IMMUNOASSAY" Plenum Publishing Corporation (Edited by T.T.Ngo), 495 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Yoshino NISHIGUCHI, Noriko TAGAWA, Fukuko WATANABE, Toshiko KIGUCHI, and Ichiya NINOMIYA: "Synthesis of 4-Carboxymethylthio-Substituted Steroids for Enzyme Immunoassay" Chem.Pharm.Bull. (Tokyo).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Yoshiharu KOBAYASHI, Yoshino NISHIGUCHI, and Fukuko WATANABE: "A Highly Sensitive Fluorescence Polarization Immunoassay by Laser Excitation" Japanese Journal of Clinical Chemistry. (Osaka).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Fukuko WATANABE: Nonisotopic Immunoassay ---Fluorescence Polarization Immunoassay---. Plenum Publishing Corporation (Edited by T.T.Ngo), 199-209 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Yoshino NISHIGUCHI: "synthesis of 4-Carboxymethylthio-Substituted Steroids for Enzyme Immunoassay." Chem.Pharm.Bull.(Tokyo).

    • Related Report
      1989 Annual Research Report
  • [Publications] 小林吉晴: "レ-ザ-励起による蛍光偏光免疫定量法の高感度化" 臨床化学.

    • Related Report
      1989 Annual Research Report
  • [Publications] Fukuko WATANABE: "NONISOTOPIC IMMUNOASSAY" Plenum Publishing Corporation(Edited by T.T.Ngo), 495 (1988)

    • Related Report
      1989 Annual Research Report
  • [Publications] Yoshino Nishiguchi.: Chem.Pharm.Bull.

    • Related Report
      1988 Annual Research Report

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Published: 1988-03-31   Modified: 2016-04-21  

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