Selective Detection of Human Serum Albumin Using a Fused-Silica Capillary Modified with Anti-Human Serum Albumin

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  • Tsukagoshi Kazuhiko
    Department of Chemical Engineering and Materials Science, Faculty of Engineering, Doshisha University
  • Indou Hirofumi
    Department of Chemical Engineering and Materials Science, Faculty of Engineering, Doshisha University
  • Sawanoi Kaori
    Department of Chemical Engineering and Materials Science, Faculty of Engineering, Doshisha University
  • Oguni Takako
    Department of Chemical Engineering and Materials Science, Faculty of Engineering, Doshisha University
  • Nakajima Riichiro
    Department of Chemical Engineering and Materials Science, Faculty of Engineering, Doshisha University

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Abstract

A fused-silica capillary modified with anti-human serum albumin was prepared using the glutaraldehyde method after activating the inner wall with a silane coupling reagent. The antibody selectively interacted via an immune reaction with the human serum albumin, which acted as an antigen. The analytical performance of the modified capillary was examined by capillary affinity chromatography using either absorption or chemiluminescence detection. The capillary affinity chromatography required only minimal reagents and samples due to the extremely small diameter of the capillary. For chemiluminescence detection, a peroxyoxalate reagent was used and human serum albumin was labeled with fluorescamine for competitive immunoassay. After optimization of the analytical conditions, chemiluminescence detection showed a higher sensitivity than absorption detection. Furthermore, a modified capillary was also used in capillary electrophoresis with the absorption detector to allow affinity capillary electrophoresis.

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