Carbon Nanofiber-based Luminol-biotin Probe for Sensitive Chemiluminescence Detection of Protein

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  • BAJ Stefan
    Department of Chemical Organic Technology and Petrochemistry, Faculty of Chemistry, Silesian University of Technology
  • KRAWCZYK Tomasz
    Department of Chemical Organic Technology and Petrochemistry, Faculty of Chemistry, Silesian University of Technology
  • PRADEL Natalia
    Department of Chemical Organic Technology and Petrochemistry, Faculty of Chemistry, Silesian University of Technology
  • 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
  • DRAGUSHA Shpend
    Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University
  • SKUTIL Krzysztof
    Department of Chemical Organic Technology and Petrochemistry, Faculty of Chemistry, Silesian University of Technology
  • PAWLYTA Miroslawa
    Institute of Engineering Materials and Biomaterials, Silesian University of Technology
  • KAI Masaaki
    Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University

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Description

A carbon nanofiber-based luminol-biotin probe was synthesized for the sensitive chemiluminescence (CL) detection of a target protein by grafting luminol and biotin onto an oxidized carbon nanofiber. This carbon nanofiber was prepared by chemical vapor-deposition with methane in the presence of the Ni–Cu–MgO catalyst, which was followed by oxidization with HNO3–H2SO4 to produce a carboxyl group on the surface of the nanofiber. The material was grafted with luminol and biotin by means of a standard carbodiimide activation of COOH groups to produce corresponding amides. The substance was water-soluble and thus could be utilized as a sensitive CL probe for a protein assay. The probe showed highly specific affinity towards the biotin-labeled antibody via a streptavidin–biotin interaction. The detection limit for this model assay was approximately 0.2 pmol of the biotinized IgG spotted on a polyvinylidene fluoride (PVDF) membrane. Nonspecific binding to other proteins was not observed. Therefore, the synthesized carbon nanofiber-based CL probe may be useful for a sensitive and specific analysis of the target protein.

Journal

  • Analytical Sciences

    Analytical Sciences 30 (11), 1051-1056, 2014

    The Japan Society for Analytical Chemistry

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