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Fabrication of Biosensors with the use of Electrodes Immobilizing Plural Kinds of Enzymes

Research Project

Project/Area Number 10555305
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 工業物理化学
Research InstitutionOsaka University

Principal Investigator

KUWABATA Susumu  Engineering, Osaka University, Associate Professor, 大学院・工学研究科, 助教授 (40186565)

Co-Investigator(Kenkyū-buntansha) YONEYAMA Hiroshi  Engineering, Osaka University, Professor, 大学院・工学研究科, 教授 (80029082)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 1999: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1998: ¥3,700,000 (Direct Cost: ¥3,700,000)
Keywordsbiosensor / cholesterol sensor / uric acid sensor / cholesterol oxidase / uricase / electron mediator / artificial bilayer / self-assembled monolayer / コレストロールエステラーゼ / コレステロールエステラーゼ
Research Abstract

(1) Fabrication of Cholesterol Sensors using Au electrodes immobilizing Cholesterol Oxidase and Cholesterol Esterase
Chemical adsorption of aminoethanethiol on a gold electrode provided its densely packed self-assembled monolayer. Cholesterol oxidase and cholesterol esterase were covalently immobilized on the electrode with use of glutaraldehyde as a cross-linking agent. The resulting electrode was polarized at 0 V vs. SCE in an electrolyte solution containing thionin as an electron mediator. It was found that injection of free cholesterol or cholesterol esterase gave generation of oxidation currents and the value increased steadily with an increase in the substrate. From these experimental results, we established a practical methodology of precise determination of concentration of free cholesterol and cholesterol esterase with the use of the prepared cholesterol sensors.
(2) Fabrication of Uric Acid Sensors using Au electrodes immobilizing Uricase and Electron Mediator
An Au electrode immobilizing uricase (uric acid oxidase) was also prepared by the same method as that mentioned above. The electrode worked as a uric acid sensor if an electrolyte solution containing an appropriate electron mediator was used. Preparation of Au electrode on which uricase and electron mediator were immobilized was achieved by using a redox polymer which can work as an electron mediator for uricase. In this case, concentration of uric acid in solution was able to be determined without addition of any other substances in the solution. Furthermore, an artificial bilayer formed by self-organized fusion of a self-assembled monolayer of alkanethiol and phospholipid was used to immobilize uricase that is one of membrane proteins together with electron mediators. The bilayer electrodes worked well as the uric acid sensor possessing an extremely high substrate selectivity.

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Susumu Kuwabata: "Preparation and properties of amperometric uric acid sensors"Sensors and Actuators B. 52. 72-77 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takahiro Nakaminami: "Amperometric determination of total cholestrol at gold electrodes covalently modified with cholestrol oxidase and cholestrol esterase with use of thionin as an electron mediator"Analytical Chemistry. 71(5). 1068-1076 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takahiro Nakaminami: "Uricase-catalyzed oxidation of uric acid usin an artificial electron acceptor and fabrication of amperometric uric acid sensors with use of a redox ladder polymer"Analytical Chemistry. 71(10). 1928-1934 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takahiro Nakaminami: "A biomimetic phospholipid/alkanethiolate bilayer immobilizing uricase and an electron mediator on on an Au electrode for amperometire determination of uric acid"Analytical Chemistry. 71(19). 4278-4283 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Susumu Kuwabata, Takahiro Nakaminami, Shin-ichiro Ito and Hiroshi Yoneyama: "Preparation and properties of amperometric uric acid sensors"Sens. Actuators, B. B52(1-2). 72-77 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takahiro Nakaminami, Shin-ichiro Ito Susumu Kuwabata and Hiroshi Yoneyama: "Amperometric determination of total cholesterol at gold electrodes covalently modified with cholesterol oxidase and cholesterol esterase with use of thionin as an electron mediator"Anal. Chem. 71(5). 1068-1076 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takahiro Nakaminami, Shin-ichiro Ito, Susumu Kuwabata and Hiroshi Yoneyama: "Uricase-catalyzed oxidation of uric acid using an artificial electron acceptor and fabrication of amperometric uric acid sensors with use of a redox ladder polymer"Anal. Chem.. 71(10). 1928-1934 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takahiro Nakaminami, Shin-ichiro Ito, Susumu Kuwabata and Hiroshi Yoneyama: "A biomimetic phospholipid/alkanethiolate bilayer immobilizing uricase and an electron mediator on an Au electrode for amperometric determination of uric acid"Anal. Chem.. 71(19). 4278-4283 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takahiro Nakaminami: "Amperometric determination of total cholesterol at gold electrodes covalently modified with cholesterol oxidase and cholesterol esterase with use of thionin as an electron mediator"Analytical Chemistry. 71(5). 1068-1076 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Takahiro Nakaminami: "Uricase-catalyzed oxidation of uric acid using an artificial electron acceptor and fabrication of amperometric uric acid sensors with use of a redox ladder polymer"Analytical Chemistry. 71(10). 1928-1934 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Takahiro Nakaminami: "A biomimetic phospholipid/alkanethiolate bilayer immobilizing uricase and an electron mediator on an Au electrode for amperometirc determination of uric acid"Analytical Chemistry. 71(19). 4278-4283 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Susumu Kuwabata: "Preparation and properties of amperometric uric acid sensors" Sensors and Actuators B. 52. 72-77 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takahiro Nakaminami: "Amperometric determination of total cholesterol at gold electrodes covalently modified with cholesterol oxidase and cholesterol esterase with use of thionin as an electron mediator" Analytical Chemistry. (印刷中). (1999)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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