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Development of Automatic Method for Determining Enzyme Activities in Serum Using Bioreactor System

Research Project

Project/Area Number 62870108
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field Laboratory medicine
Research InstitutionKyoto University

Principal Investigator

MURACHI Takashi  Faculty of Medicine, Kyoto University, 医学部, 教授 (10089104)

Co-Investigator(Kenkyū-buntansha) TOTANI Masayuki  Faculty of Medicine, Kyoto University, 医学部, 講師 (70163988)
TABATA Masayoshi  College of Medical Technology, Kyoto University, 医療技術短期大学部, 助教授 (70115880)
Project Period (FY) 1987 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥16,000,000 (Direct Cost: ¥16,000,000)
Fiscal Year 1988: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1987: ¥13,000,000 (Direct Cost: ¥13,000,000)
Keywordsbioreactor / FIA / chemiluminometry / serum enzyme activity / lactate dehydrogenase (LDH) / 血清酵素活性 / 乳酸脱水素酵素(LDH)
Research Abstract

Enzyme activities in serum have been determined widely using an automated analyzer in clinical analysis. Recently, the automated analysis of nonprotein organic compounds in serum by a bioreactor with immobilized wnzyme in column form has been put to practical use. For the purpose of applying the bioreactor to the determination of enzyme activities in serum and increasing the range of its use, we studied as follows, and obtained a lot of results.
(1) Study of the incubation system for the reaction of enzyme. Since the system putting a reaction coil for the reaction of enzyme as a bypath in the flow injection analysis (FIA) system was simple and easier to remove sample blanks, we chose the system.
(2) Development of the automatic analysis of enzyme activities in serum using a bioreactor. 1. Determination of lactate dehydrogenase (LDH) activities in serum. LDH reaction was taken place using pyruvate as the substrate in the reaction coil put as a bypath in the FIA system, and lactate produced by LDH reaction was oxidized to pyruvate and hydrogen peroxide by the lactate oxidase bioreactor. Hydrogen peroxide produced was determined by chemiluminescence method. Endogenous lactate was removed by lactate oxidase・catalase precolumn. The results obtained were excellent, and correlated satisfactorily with those by a Hitachi Model 726 automatic analyzer. The present method did not need any sample blank.
2. Determination of amylase activity in serum. Maltopentaose was used as the substrate for amylase reaction. Maltose produced by amylase reaction was catalyzed to hydrogen peroxide by the maltose phosphorylase・pyranose oxidase bioreactor. Glucokinase column was used to remove endogenous glucose in serum. The results obtained was good.

Report

(2 results)
  • 1988 Annual Research Report   Final Research Report Summary
  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] 田畑勝好: 化学と生物. 25. 425-427 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Matashige Ohyabu: Analytical Sciences. 3. 277-278 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Takashi Murachi: Biotechnolo gy and Applied Biochemistry. 9. 303-309 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Masayoshi Tabata: Annals of Clinlcal Biochemistry. (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Masayoshi Tabata: Journal of Bioluminescence and Chemiluminescence. 2. 63-67 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Masayoshi Tabata: Journal of Bioluminescence and Chemiluminescence. (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 村地孝: "標準臨床検査医学、分担課題「血清酵素と尿酵素」" 医学書院, 16 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 田畑勝好、村地孝: "生物発光と化学発光(基礎と実験)、「化学発光を利用した臨床化学分析」「臨床化学分析への応用」" 廣川書店, 24 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Matashige,Ohyabu: ""One-Shot" flow injection analysis with immobilized enzyme column: clinical application" Analytical Sciences. 3. 277-278 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Takashi.Murachi: "Use of a bioreactor consisting of sequentially aligned L-glutamate dehydrogenase and L-glutamate oxidase for the determination of ammonia by chemiluminescence" Biotechnology and Applied Biochemistry. 9. 303-309 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Masayoshi,Tabata: "Determination of lactic acid and LDH activity on FIA system using immobilized enzyme column and chemiluminescence" Annals of Clinical Biochemistry. in press. (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Masayoshi,Tabata: "A chemiluminometric method for the determination of urea in serum using a three-enzyme bioreactor" Journal of Bioluminescence and Chemiluminescence. 2. 63-67 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Masayoshi,Tabata: "A chemiluminescence automatic analyzer for the measurement of biological compounds" Journal of Bioluminescence and Chemiluminescence. (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Takashi,Murachi: GBF Monographs Vol. 10 Bioreactor system for clinical analysis. VCH Verlagsgesellschaft, Weinheim, Jermany, 119-124 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Masayoshi Tabata: Journal of Bioluminescence and Chemiluminescence. 2. 63-67 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Masayoshi Tabata: Journal of Bioluminescence and Chemiluminescence. (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Masayoshi Tabata: Biomedical Chromatography.

    • Related Report
      1988 Annual Research Report
  • [Publications] Masayoshi Tabata: Annals of Clinical Biochemistry.

    • Related Report
      1988 Annual Research Report
  • [Publications] 田畑勝好,村地孝: "生物発光と化学発光(基礎と実験)「化学発光を利用した臨床化学分析」「臨床化学分析への応用」" 廣川書店, 231-247,270-276 (1988)

    • Related Report
      1988 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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