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Simultaneous Determination of Cyc and Vegf165 Tumor Markers Based on Immobilization of Flavin Adenine Dinucleotide and Thionine As Probes on Reduced Graphene Oxide-Poly(Amidoamine)/Gold Nanocomposite Modified Dual Working Screen-Printed Electrode Publisher



Amouzadeh Tabrizi M1 ; Shamsipur M2 ; Saber R1, 3 ; Sarkar S4
Authors
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Authors Affiliations
  1. 1. Research Center for Science and Technology in Medicine, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Department of Chemistry, Razi University, Kermanshah, Iran
  3. 3. School of Advanced Technologies in Medicine Sciences, Tehran University of Medical, Iran
  4. 4. Department of Medical Physics and Biomedical Engineering, Tehran University of Medical Sciences, Tehran, Iran

Source: Sensors and Actuators# B: Chemical Published:2017


Abstract

Herein, a novel electrochemical bi-aptasensor using green synthesized reduced graphene-poly(amidoamine)/gold nanocomposite (rGO-PAMAM/Aunano) as supporting matrix for covalent immobilization of flavin adenine dinucleotide (FAD) and thionine (Th) on the first (W1) and second (W2) surfaces of dual working electrode screen printed electrode has proposed. FAD and Th probes were immobilized by glutaraldehyde linker based on a reaction between primary amines (–NH2) of FAD and Th with –NH2 of rGO-PAMAM/Aunano modified dual working electrode for simultaneous determination of cytochrome c (CYC) and vascular endothelial growth factor (VEGF165) tumor markers. The principle response of proposed bi-aptasensor was based on the selective interaction of CYC with W1/rGO-PAMAM-FAD/Aunano/Anti-ptamerCYC and VEGF165 with W2/rGO-PAMAM-Th/Aunano/Anti-ptamerVEGF165, respectively. This selective interaction, leads to decrease of response currents of immobilized electrochemical probes of FAD and Th that have different formal potential (E0′). Electrochemical behavior of bi-aptasensor was studied by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The liner calibration curve for tumor markers determination was 2.5–320.0 pM. The limit of detections (LOD) for CYC was 1.0 pM and for VEGF165 was 0.7 pM (3σ/S). The biomarker-aptamer dissociation constant (kd) was calculated based on the change in current to be 63.9 pM for CYC/Anti-aptamerCYC and 38.4 pM for VEGF165/Anti-aptamerVEGF165, respectively. This bi-aptasensor was used for the determination of VEGF165 and CYC in the human serum sample. © 2016
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