Selectivity and Regeneratability Evaluation of Antigen Analogous-Modified Surface Film for Polychlorinated Dibenzodioxin Immunosensor Using Surface Plasmon Resonance Measurement

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  • YASUZAWA Mikito
    Department of Chemical Science and Technology, Institute of Technology and Science, The University of Tokushima
  • FUTAGAMI Norimichi
    Department of Chemical Science and Technology, Graduate School of Advanced Technology and Science, The University of Tokushima
  • FUCHIWAKI Yusuke
    Department of Chemical Science and Technology, Graduate School of Advanced Technology and Science, The University of Tokushima
  • YABUTANI Tomoki
    Department of Chemical Science and Technology, Institute of Technology and Science, The University of Tokushima
  • MOTONAKA Junko
    Department of Chemical Science and Technology, Institute of Technology and Science, The University of Tokushima

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Other Title
  • ダイオキシン計測のための表面プラズモンイムノセンサー用感応膜の試作と評価
  • ダイオキシン ケイソク ノ タメ ノ ヒョウメン プラズモンイムノセンサーヨウ カンノウ マク ノ シサク ト ヒョウカ

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Abstract

For the development of a sensitive and accurate immunosensor, the production of a highly specific surface to the corresponding antibody is essential. Moreover, a specific surface having good reusability is worthwhile for constructing a prompt auto-monitoring immunosensing system. In this study, antigen analogous-modified gold substrates having a specific adsorption property to an anti-dioxin antibody, were prepared by the immobilization of 2,4,5-trichlorophenoxy (TCP) derivatives on a gold surface using various methods. An anti-dioxin antibody, which has a good correlation immunoaffinity with the total toxic equivalent quantity (TEQ) of polychlorinated dibenzo-p-dioxins, was employed as a specific antibody, and the specification of the surface was evaluated by a comparison of the surface plasmon resonance (SPR) responses to a specific anti-dioxin antibody and non-specific anti-hemoglobin antibody. In order to obtain a firm and stable immobilization of TCP, a carboxyl group was introduced on gold by a self-assemble monolayer (SAM) technique, and TCP derivatives were attached to the SAM surface with covalent bonding directly and indirectly through the presence of a hexamethylene chain, bovine serum albumin (BSA), and ethylene glycol oligomer (EGO). A TCP derivative-immobilized substrate, prepared using BSA, showed poor reusability. A TCP derivative-immobilized substrate having EGO was highly specific to an anti-dioxin antibody, while that prepared by the direct immobilization of TCP and with introduction of a hexamethylene chain presented nonspecific adsorption. In addition, a TCP derivative-immobilized substrate having EGO showed a good reproducible response from the second repetition, and was reusable more than seven times.

Journal

  • BUNSEKI KAGAKU

    BUNSEKI KAGAKU 60 (1), 81-86, 2011

    The Japan Society for Analytical Chemistry

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