Monitoring of troponin release from cardiomyocytes during exposure to toxic substances using surface plasmon resonance biosensing

Abstract

Troponin T (TnT) is a useful biomarker for studying drug-induced toxicity effects on cardiac cells. In this paper, 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 for analysing of the TnT release. The detection limit of TnT was determined to 30 ng/ml in a direct assay set-up and to 10 ng/ml in a sandwich assay format. Exposure of doxorubicin, troglitazone, quinidine and cobalt chloride to the cardiomyocytes for periods of 6 to 24 hours gave significant SPR responses, while 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.

Orientation and capturing of antibody affinity ligands: Applications to surface plasmon resonance biochips

“…Human serum albumin is a common model protein and also a critical biomarker for hepatocyte differentiation and function often used in vitro [23,24]. Troponin T is an important clinical biomarker for myocardial states, and is one of the endpoints for in vitro cardiotoxicity testing [25,26].…”

Mesenchymal Stem Cells: An Innovative Approach in Pharmacokinetics

“…This new approach utilizes a dual-channel automated image cytometer that allows for simultaneous measurement of the cardiomyocyte action potential and calcium transient using voltage and calcium-sensitive dyes. ESC-and iPSCderived cardiomyocytes have recently been used to study doxorubicintriggered toxicity [62]. MSCs-derived cardiomyocytes are suitable to study the effects of compounds which do not interfere with the ion channel functions but still cause cardiotoxicity.…”

“…Troponin T (TnT) is a useful biomarker for studying drug-induced toxicity effects on cardiac cells. After induction of doxorubicin, MSCs-derived cardiomyocytes released detectable levels of cardiac TnT and fatty acid-binding protein 3 in a dose-dependent manner [62]. Based on the availability of very sensitive and rapid analytical tools for these biomarkers, the assay lends itself well to miniaturization and high-throughput formats.…”

Drug Discovery Models and Toxicity Testing Using Embryonic and Induced Pluripotent Stem-Cell-Derived Cardiac and Neuronal Cells

“…Two clinically decisive biomarkers of cardiac damage that are sensitive indicators for doxorubicin-induced toxicity were studied. The ESC- and iPSC-derived cardiomyocytes released detectable levels of cardiac troponin T and fatty acid-binding protein 3 in a dose-dependent manner after doxorubicin induction [82]. Based on the availability of very sensitive and rapid analytical tools for these biomarkers, the assay lends itself well to miniaturization and high-throughput formats.…”