Sensitive Chemiluminescence Detection of Prion Protein on a Membrane by Using a Peroxidase-Labeled Dextran Probe

• AZAM Md. Golam

  Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University

• SHIBATA Takayuki

  Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University

• KABASHIMA Tsutomu

  Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University

• KAI Masaaki

  Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University Global COE Program, Nagasaki University

Abstract

We synthesized dextran-based macromolecular probes carrying multiple molecules of horseradish peroxidase (HRP) as a signal-trigger enzyme and of biotin as an assembly mediator without losing the enzymatic activity. Multiple attachments of HRP and biotin to the dextran backbone were readily accomplished after the formation of aldehyde groups into the dextran macromolecule by periodate oxidation. The synthesized macromolecular probe was successfully used for sensitive chemiluminescence (CL)-imaging detection of mouse recombinant prion protein on a nylon membrane. The prion protein at a small amount of 20 fmol blotted on a nylon membrane was specifically detected, indicating at least a 10-times higher sensitivity than that of a conventional biotinylated HRP probe. Therefore, the synthesized dextran-based probes containing HRP and biotin should be used for the sensitive high-throughput analysis of various proteins on a solid-phase membrane.

Journal

• Analytical Sciences

  Analytical Sciences 27 (7), 715-, 2011

  The Japan Society for Analytical Chemistry

Keywords

• Analytical Chemistry

Details

• CRID: 1390001204261094528
• NII Article ID: 10029332555
• NII Book ID: AA10500785
• DOI: 10.2116/analsci.27.715
• COI: 1:STN:280:DC%2BC3MnntlWjuw%3D%3D
• ISSN: 13482246
• ISSN: 09106340
• HANDLE: 10069/25734
• NDL BIB ID: 11154238
• PubMed: 21747179
• Web Site: https://nagasaki-u.repo.nii.ac.jp/records/13655
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I11154238
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.27.715.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.27.715/fulltext.html
• Web Site: http://www.jstage.jst.go.jp/article/analsci/27/7/27_7_715/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2012093686
• Text Lang: en
• Data Source:
  • JaLC
  • IRDB
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed