Development of an optical immunosensor based on the fluorescence of Cyanine-5 for veterinarian diagnostics |
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Authors: | Marta Silva Helder Cruz Osvaldo Rossetti Alicia Arese Abel Oliva |
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Institution: | Biosensors Laboratory, IBET/ITQB- Instituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Química e Biológica, Apartado 12, P-2781-901 Oeiras, Portugal. |
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Abstract: | A model optical immunosensor was developed to quantify an antibody present in a sample by measuring the fluorescence of Cyanine-5 conjugated with the antibody, using a competitive and a sandwich immunoreaction configuration, with the antigen immobilised in controlled pore glass beads. At pH 2, 94% of the antigen-antibody complex was dissociated, allowing reutilisation. Photobleaching had no effect on the fluorescence. This model system was used to detect Brucella sp. infection and could quantify anti-Brucella sp. antibodies in ovine serum samples in the range from 0.005 to 0.11 mg ml(-1). |
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Keywords: | Brucella sp diagnostics controlled pore glass immobilization optical biosensor photobleaching |
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