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Enhanced Electrochemiluminescence Biosensing of Gene-Specific Methylation in Thyroid Cancer Patients' Plasma Based Integrated Graphitic Carbon Nitride-Encapsulated Metal-Organic Framework Nanozyme Optimized by Central Composite Design Publisher



Khoshfetrat SM1, 2 ; Dorraji PS3 ; Fotouhi L3, 4 ; Hosseini M1 ; Khatami F5 ; Moazami HR6 ; Omidfar K2, 7
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Authors Affiliations
  1. 1. Department of Chemistry, Faculty of Basic Science, Ayatollah Boroujerdi University, Boroujerd, Iran
  2. 2. Biosensor Research Center, Endocrinology and Metabolism Molecular–Cellular Sciences Institute, Tehran University of Medical Sciences, P.O. Box 14395/1179, Tehran, Iran
  3. 3. Department of Chemistry, Faculty of Physics and Chemistry, Alzahra University, Tehran, Iran
  4. 4. Analytical and Bioanalytical Research Centre (ABRC), Alzahra University, Tehran, Iran
  5. 5. Urology Research Center, Tehran University of Medical Sciences, Tehran, Iran
  6. 6. Nuclear Fuel Cycle Research School, Nuclear Science and Technology Research Institute, P.O. Box: 11365–8486, Tehran, Iran
  7. 7. Endocrinology and Metabolism Research Center, Endocrinology and Metabolism Research Institute, Tehran University of Medical Sciences, Tehran, Iran

Source: Sensors and Actuators B: Chemical Published:2022


Abstract

Circulating cell free DNA (cfDNA) methylation is a novel type of cancer biomarker, but its minuscule proportion of total DNA makes proper analysis difficult in clinical samples. Herein, a sensitive electrochemiluminescence immuno-DNA sensor was designed to analyze DNA methylation using sandwiching the target methylated DNA between the magnetic nanoparticles/anti-5-methylcytosine monoclonal antibody (MNPs/anti-5mc) bioconjugate and luminol-loaded within phosphorylated DNA capture probe-immobilized C3N4 NS@UiO-66 core@shell nanozyme. Taking advantages of increased concentration of C3N4 NS nanozymes' •OH-generation, nanoproximity effect of C3N4 NS and luminol, high density coordination of capture probe on the UiO-66 metal organic framework (MOF), MNPs' function in improving the signal-to-background ratio (S/B) in complicated plasma media, and remarkable electrocatalytic activity of reduced graphene oxide-modified pencil graphite electrode (rGO/PGE), multiple signal amplification was achieved without bisulfite and PCR amplification. The immuno-DNA sensor offers a linear response across a wide dynamic range from 20 pg to 20 ng, with a detection limit of 10 pg, when optimized by a face-centered central composite design (FCCD). Our method can differentiate methylation levels as low as 0.1%. Tumor-specific methylation DNA is definitely identified in the plasma of 9 of 10 thyroid cancer patients' plasma. The 91% clinical sensitivity implies strong clinical diagnosis consistency. The suggested method was successfully utilized to evaluate methylated DNA in human blood plasma, demonstrating the platform's potential for disease diagnostics and biochemistry research. © 2022 Elsevier B.V.
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