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Monitoring of troponin release from cardiomyocytes during exposure to toxic substances using surface plasmon resonance biosensing
Authors:Henrik Andersson  Bertil Kågedal  Carl-Fredrik Mandenius
Institution:1. Division of Biotechnology/IFM, Link?ping University, 581 83, Link?ping, Sweden
2. Division of Clinical Chemistry, Department of Clinical and Experimental Medicine, Link?ping University, 581 83, Link?ping, Sweden
Abstract:Troponin T (TnT) is a useful biomarker for studying drug-induced toxicity effects on cardiac cells. We describe how a surface plasmon resonance (SPR) biosensor was applied to monitor the release of TnT from active HL-1 cardiomyocytes in vitro when exposed to cardiotoxic substances. Two monoclonal human TnT antibodies were compared in the SPR immunosensor to analyse the TnT release. The detection limit of TnT was determined to be 30 ng/ml in a direct assay set-up and to be 10 ng/ml in a sandwich assay format. Exposure of the cardiomyocytes to doxorubicin, troglitazone, quinidine and cobalt chloride for periods of 6 and 24 h gave significant SPR responses, whereas substances with low toxicity showed insignificant effects (ascorbic acid, methotrexate). The SPR results were verified with a validated immunochemiluminescence method which showed a correlation of r 2 = 0.790.
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