β-Lactamase label-based potentiometric biosensor for α-2 interferon detection |
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Authors: | T. A. Sergeyeva, A. P. Soldatkin, A. E. Rachkov, M. I. Tereschenko, S. A. Piletsky,A. V. El skaya |
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Affiliation: | T. A. Sergeyeva, A. P. Soldatkin, A. E. Rachkov, M. I. Tereschenko, S. A. Piletsky,A. V. El‘skaya |
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Abstract: | An effective immunosensor for α-2 interferon detection based on pH-sensitive field effect transistor (pH-FET) has been developed. A specific sensing element was fabricated by immobilizing α-2 interferon on the gate of a pH-FET. The interaction of anti-interferon antibodies labelled with β-lactamase with interferon–pH-FET (in the presence of specific enzyme substrate) leads to a local pH-change at the surface of transducer and produces an electrochemical signal which is proportional to the conjugate concentration. The main performance characteristics of the sensor obtained (sensitivity, dynamic range, operational and storage stability) were estimated. The effect of pH, buffer concentration and ionic strength on the immunosensor response as well as conditions of immunosensor regeneration were studied. For the determination of the interferon concentration in a sample solution the competitive electrochemical immunoassay has been employed. A linear response of the sensor on α-2 interferon concentration is obtained in the range 10–100 μg/ml. This gives the possibility to detect α-2 interferon in a non-diluted cultivated broth. The data of the competitive electrochemical immunoassay are available within 30 min and are in good accordance with the enzyme-linked immunosorbent assays. |
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Keywords: | β -Lactamase α -2 Interferon pH-sensitive field effect transistors Biosensor Electrochemical immunoassay |
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