Protein functionalized nanostructured zirconia based electrochemical immunosensor for cardiac troponin I detection
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We report results of the studies relating to the fabrication of nanostructured zirconia (nZrO2) based immunosensor for cardiac troponin I biomarker (acute myocardial infarction) detection. One step, low temperature hydrothermal process was used for the synthesis of nZrO2 (;5 nm). This nZrO2 was functionalized with 3-aminopropyl triethoxy silane (APTES) and thereafter it was electrophoretically deposited on to indium tin oxide (ITO) coated glass electrode. EDC/NHS surface chemistry was used for covalent immobilization of monoclonal anti-troponin-I (anti-cTnI) antibodies onto APTES/nZrO2/ITO electrode. The structural, morphological and functional characterization of the synthesized nanoparticles and the fabricated immunoelectrode were conducted via X-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), and electrochemical techniques. The results of electrochemical response studies of BSA/anti-cTnI/APTES/nZrO2/ITO immunoelectrode reveal that, this platform can be used for efficient detection of cardiac troponin I (cTnI) biomarker with a wide linear detection range (0.1–100 ng/mL) and sensitivity [3.9 lA mL/(ng cm2)].
I. INTRODUCTION
Among the various cardiovascular diseases, acute myocardial infarction (AMI) is currently one of the leading cause of death worldwide.1,2 The main symptoms associated with AMI are chest pain, shortness of breath, nausea, heartburn etc. that occur due to the blocking of blood flow in the circulatory system.3 High level of cholesterol present in blood leads to the blockage of coronary artery resulting in damage of cardiac muscles.2,3 Troponin is an integral protein present in each myofibril and helps in contraction of the cardiac muscles.4 It is a complex of three regulatory proteins: troponin C, troponin T, and troponin I.4 Among these, cardiac troponin I (cTnI) is released in the bloodstream when cardiac muscle is damaged or dead.5 Therefore, cTnI is presently a gold standard for the confirmation of cardiac tissue injury and can be used as an early diagnosis of AMI.6 cTnI is a water soluble proteinaceous biomarker having molecular weight of 21 kDa and is secreted in blood serum.7 The cutoff concentration of cTnI protein in the blood serum is found to be in the range, 0.01–0.1 ng/ mL and increases upto 100 ng/mL in a heart patient.5 The initial time of cTnI elevation in blood has been found to be 4–6 h and returns to normal level in 6–8 days.5 For the detection of cTnI many immunoassay techniques such as Contributing Editor: Venkatesan Renugopalakrishnan a) Address all correspondence to this author. e-mail: [email protected] DOI: 10.1557/jmr.2017.102
radioactive immune assay (RIA), fluoro-immuno assay, immuno-electrophoresis, spectro-photometric methods, electrochemiluminescence assays, radioimmunoassay, enzyme linked immunosorbent assay (ELISA) etc. can be used.8–11 These methods are time-consuming, expensive, and require highly skilled personnel. In this context, biosensors offer a simple reliable and user friendly detection s
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