Direct Determination of Horseradish Peroxidase Encapsulated in Liposomes by Using Luminol Chemiluminescence

• KAMIDATE Tamio

  Division of Biotechnology and Molecular Chemistry, Graduate School of Engineering, Hokkaido University

• KOMATSU Kanako

  Division of Biotechnology and Molecular Chemistry, Graduate School of Engineering, Hokkaido University

• TANI Hirofumi

  Division of Biotechnology and Molecular Chemistry, Graduate School of Engineering, Hokkaido University

• ISHIDA Akihiko

  Division of Biotechnology and Molecular Chemistry, Graduate School of Engineering, Hokkaido University

Abstract

Horseradish peroxidase (HRP) encapsulated in liposomes was directly detected by using luminol chemiluminescence (CL) with H₂O₂ without lysis of liposomes. At a low concentration of H₂O₂, the initial rate of HRP-catalyzed luminol CL in liposomes was slower than that of HRP-catalyzed luminol CL in a lipid-free bulk solution. The decrease in the initial rate of the CL reaction in liposomes was due to the membrane permeation of luminol and H₂O₂. At a high concentration of H₂O₂, the initial rate of the CL reaction in liposomes was the same as that in a lipid-free bulk solution. The CL measurement conditions in both a lipid-free bulk solution and in liposomes were optimized in the concentrations of luminol and H₂O₂ by measuring the CL response curves, in which only one peak appeared and the CL intensity was maximal. The CL intensity observed in HRP-catalyzed luminol CL in liposomes was a factor of seven greater than that observed in a lipid-free bulk solution. The CL intensity was dependent on the amount of HRP-encapsulated liposomes used. The detection limit in the direct detection of HRP encapsulated in liposomes was sensitive by a factor of 3 compared with that in HRP-catalyzed luminol CL in a lipid-free bulk solution.

Journal

• Analytical Sciences

  Analytical Sciences 24 (4), 477-481, 2008

  The Japan Society for Analytical Chemistry

Keywords

• Horseradish peroxidase (HRP)
• chemiluminescence (CL)

Details

• CRID: 1390001204256200192
• NII Article ID: 10021099544
• NII Book ID: AA10500785
• DOI: 10.2116/analsci.24.477
• ISSN: 13482246
• ISSN: 09106340
• HANDLE: 2115/71700
• NDL BIB ID: 9452085
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I9452085
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.24.477.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.24.477/fulltext.html
• Web Site: https://www.jstage.jst.go.jp/article/analsci/24/4/24_4_477/_pdf
• Text Lang: en
• Data Source:
  • JaLC
  • IRDB
  • NDL
  • Crossref
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed