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Development of new method for the preparation of enzyme thin films

Research Project

Project/Area Number 09558110
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Biomedical engineering/Biological material science
Research InstitutionTOHOKU UNIVERSITY

Principal Investigator

ANZAI Jun-ichi  Grad. School Pharm. Sci., Tohoku Univ., Professor, 大学院・薬学研究科, 教授 (40159520)

Co-Investigator(Kenkyū-buntansha) HOSHI Tomonori  Grad. School Pharm. Sci., Tohoku Univ., Instructor, 大学院・薬学研究科, 助手 (50302170)
KASHIWAGI Yoshitomo  Grad. School Pharm. Sci., Tohoku Univ., Instructor, 大学院・薬学研究科, 助手 (50204384)
SUZUKI Iwao  Grad. School Pharm. Sci., Tohoku Univ., Assistant Professor, 大学院・薬学研究科, 講師 (30226493)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥13,500,000 (Direct Cost: ¥13,500,000)
Fiscal Year 1999: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1998: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1997: ¥6,800,000 (Direct Cost: ¥6,800,000)
Keywordsimmobilized enzyme / enzyme film / biosensor / thin film / durability / avidin / biotin / 酵素累積膜 / 酵素センサー / 触媒活性 / 酵素 / 合成ポリマー
Research Abstract

Experimental variables for the preparation of enzyme thin films using avidin-biotin technology were investigated in detail. The enzyme thin films containing glucose oxidase (GOx) were found to be used for several months if the GOx films were prepared under the optimum conditions, including the concentration of enzyme solution, temperature, pH, and treatment time. In order to immobilize much amount of enzymes in durable thin films, some kinds of synthetic polymer were employed as material for the film. Biotin-modified polyamines (polyethyleneimine (PEI) and polyallylamine (PAA)) can be used for immobilizing much amount of avidin through avidin-biotin interactions. On the other hand, polyanions such as polyvinylsulfate (PVS) and polystyrenesulfonate (PSS) were useful for immobilize avidin through electrostatic force of attraction, by a layer-by-layer deposition of the polyanion and avidin. The flexible backbone of PVS and PSS is responsible for high loading of avidin in the thin films. In these polyanion/avidin thin films, the biological activity of avidin (or binding ability to biotin) was retained and it was possible to accommodate some biotin-tagged functional molecules in the polyanion/avidin thin films. During the preparation of enzyme thin films by avidin-biotin interactions, it necessary to keep the film wet for the development of well-organized and biologically-active films. If the film was dried, the catalytic activity of enzyme was degraded rapidly, probably due to the degradation of the structure of the film. During the course of this study, we obtained useful information for the development of biologically active enzyme thin films through avidin-biotin interactions.

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Anzai Jun-ichi: "Enzyme Multilayer-Modified Biosensors. Use of Streptavidin and Deglycosylated Avidin for Constructing Glucose Oxidase and Lactate Oxidase Multilayers"Analytical Sciences. 13. 859-861 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Anzai Jun-ichi: "Layer-by-layer Deposition of Avidin and Polymers on a Solid Surface to Prepare Thin Films : Significant Effects of Molecular Geometry of the Polymers on the Deposition Behavior"Journal of Chemical Society, Perkin Transactions 2. 1887-1889 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Anzai Jun-ichi: "Layer-by-layer Construction of Enzyme Multilayers on an Electrode for the Preparation of Glucose and Lactate Sensors : Elimination of Ascorbate Interference by Means of an Ascorbate Oxidase Multilayers"Analytical Chemistry. 70. 811-817 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Anzai Jun-ichi: "Enzyme Sensors Prepared by Layer-by-layer Deposition of Enzymes on a Platinum Electrode through Avidin-Biotin Interaction"Sensors & Actuators B. 52. 3-9 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Anzai Jun-ichi: "Layer-by-layer Construction of Multilayer Thin Films Composed of Avidin and Biotin-labeled Poly(amine)s"Langmuir. 15. 221-226 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Anzai Jun-ichi: "Preparation of Active Avidin Films by a Alyer-bylayer Deposition of Poly(vinyl sulfate)and Avidin on a Solid Surface"Journal of Chemical Society, Perkin Transactions 2. 2413-2414 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Jun-ichi Anzai: "Enzyme Multilayer-Modified Biosensors. Use of Streptavidin and Deglycosylated Avidin for Constructing Glucose Oxidase and Lactate Oxidase Multilayers"Anal. Sci.. Vol. 13. 859-861 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Jun-ichi Anzai: "Layer-by-layer Deposition of Avidin and Polymers on a Solid Surface to Prepare Thin Films ; Significant Effects of Molecular Geometry of the Polymers on the Deposition Behavior"J. Chem. Soc.. Perkin 2. 1887-1889 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Jun-ichi Anzai: "Layer-by-layer Construction of Enzyme Multilayers on an Electrode for the Preparation of Glucose and Lactate Sensors ; Elimination of Ascorbate Interference by Means of an Ascorbate Oxidase Multilayers"Anal. Chem.. Vol. 70. 811-817 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Jun-ichi Anzai: "Enzyme Sensors Prepared by Layer-by-layer Deposition of Enzymes an a Platinum Electrode through Avidin-biotin Interaction"Sens. Actuators B. Vol. 52. 3-9 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Jun-ichi Anzai: "Layer-by-layer Construction of Multilayer Thin Films Composed of Avidin and Biotin-labeled Poly(amine)s"Langmuir. Vol. 15. 221-226 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Jun-ichi Anzai: "Preparation of Active Avidin Films by a Layer-by-layer Deposition of Poly(vinyl sulfate) and Avid on a Solid Surface"J. Chem. Soc.. Perkin 2. 2413-2414 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Anzai Jun-ichi: "Preparation of active avidin films by a layer-by-layer deposition of poly (vinyl sulfate) and avidin on a solid surface"Journal of Chemical Society,Perkin Transactions 2. 2413-2414 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Anzai,Jun-ichi: "Layer-by-layer construction of multilayer thin films composed of avidin and biotin-labeled poly(amine)s" Langmuir. 15・1. 221-226 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Anzai,Jun-ichi: "Layer-by-layer deposition of avidin and polymers on a solid surface to prepare thin films : significant effects of molecular geometry of the polymers on the deposition behaviour" J.Chem.Soc., Perkin Transaction 2. 1887-1889 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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