Competitive Immunoassay Using Capillary Electrophoresis with a Chemiluminescence Detector

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  • Tsukagoshi Kazuhiko
    Department of Chemical Engineering and Materials Science, Faculty of Engineering, Doshisha University
  • Jinno Naoya
    Department of Chemical Engineering and Materials Science, Faculty of Engineering, Doshisha University
  • Toguchi Kae
    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

We have proposed competitive immunoassay using capillary electrophoresis (CE) with a chemiluminescence (CL) detector, in which a small amount of sample (ca. 20 μL) is required for analysis. Human serum albumin (HSA) and anti-human serum IgG (anti-HSA) were used for immune reaction as a model. A luminol and hydrogen peroxide CL reaction was adopted, and HSA was labeled with isoluminol isothiocyanate (ILITC) for competitive immunoassay. The reactant after the immune reaction was directly subjected to CE with the CL detector, where the labeled HSA was easily and rapidly separated and detected. The amount of labeled HSA indicated a good relationship to that of HSA as an analyte through the immune reaction. The HSA was determined over the range of 0.2–1.2 μM with a detection limit of 0.1 μM (S/N = 3). The present method features high sensitivity, a small sample volume, and easy and rapid operation. The method also shows the possibility to analyze a specific protein in a serum sample.

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