Bioelectrochemical Sensors Prepared by Nanobiotechnological Approaches and Their Applications in Biomarker Analysis in Biological Fluids
Project no. 25271 · Status: Kapanmış (Yayın bekleniyor) · 2023
Summary
<p>Electrochemical immunosensors allow the diagnosis of diseases at an early stage thanks to their ability to detect biomarkers with high specificity and sensitivity even at very low concentrations in biological samples. Detection of disease biomarkers, especially in diagnostically rich biological fluids such as pleural fluids, allows early and effective diagnosis of many serious diseases, especially lung cancer, tuberculosis and pleural infections. Early diagnosis prevents disease progression, reduces the risk of complications and improves quality of life, as it allows earlier initiation of treatment. Therefore, the development of new generation electrochemical immunosensors and the use of these sensors in critical diagnostic samples such as pleural fluid are of great importance in terms of both improving diagnostic processes and reducing the burden on healthcare systems in modern medicine.</p><p>In this project, a new electrochemical immunosensor system was developed for the simultaneous determination of C-Reactive Protein (CRP), Mesothelin (MSLN), Carcinoembryonic Antigen (CEA) and <em>M.tuberculosis</em> (MPt64 protein) analytes in pleural fluid samples. For this purpose, firstly mannose-4 aminophenol-L-alanine (Man-APA) sugar ligands were synthesized. They were characterized by FTIR, MS, MS/MS, 1H and 1H COSY NMR techniques. Then, Man-APA was modified with AgNPs and characterized by Uv-vis, XRD and HRTEM techniques. These synthesized AgNPs/Man-4APAs were deposited on the electrode surface. Surface and elemental characterizations were performed by SEM-EDS. Then, Anti-CRP, Anti-MSLN, Anti-CEA and Anti-MPT64 antibodies were immobilized on the electrode surfaces separately by EDC-NHS carboimide chemistry. Covalent conjugation was performed between the amine groups of AgNPs/P(Man-4APA) and the carboxyl groups of the antibodies and characterized by XPS technique. Linear detection range and detection limit (LOD) were calculated for each analyte. The effect of some potential interfering compounds on the current signal was tested. Then, CRP, MSLN, CEA and MPT64 were simultaneously determined in pleural fluids from patients with pleural effusion. Finally, the data obtained after electrochemical measurements were tested for analysis with machine learning.</p>