A new strategy to design label-free electrochemical biosensor for ultrasensitive diagnosis of CYFRA 21-1 as a biomarker for detection of non-small cell lung cancer

Abstract

Lung cancer is one of the most dangerous cancers with high mortality rate among other cancers therefore, early detection of this cancer is very important. Many studies have been reported in ways of diagnostic lung cancer early. According to reports, one of the most important biomarkers to detect lung cancer is Cytokeratin 19 fragment 21-1 (CYFRA21-1), which is significantly related to non-small cell lung cancer, in particular, squamous cell carcinoma. Thus, finding a new method for the early diagnosis of CYFRA 21-1 (DNA target probe) is essential. In the present report, we design a novel label-free electrochemical DNA-biosensor related to the signal of guanine oxidation. The proposed DNA biosensor is fabricated by a modified glassy carbon electrode (GCE) with reduced-graphene oxide (rGO), poly pyrrole (PPy), silver nanoparticles (AgNPs) and single-strand DNA (ssDNA as capture probe) GCE/rGO/PPy/AgNPs/ssDNA. The differential pulse voltammetry (DPV) and cyclic voltammetry (CV) techniques are used to verify the hybridization process between capture and target probes. Electrochemical impedance spectroscopy (EIS), energy diffraction X-ray (EDX) and field-emission scanning microscopy (FE-SEM) techniques are applied to the characterization of different modified GCE surfaces as well as X-ray diffraction (XRD) for graphene oxide synthesis. The XRD pattern of the synthesized GO that its diffraction peak appears at 10.2. The applied CV and DPV for the guanine oxidation are determined under optimal conditions. The label-free DNA biosensor showed a great result for the determination of CYFRA21-1 with a wide linear range from two consecutive linear relationships of peak current and CYFRA21-1 concentration were found (1.0 × 10^-14 - 1.0 × 10^-10 M, R² = 0.9936 and 1.0 × 10^-9 - 1.0 × 10^-6 M, R² = 0.9955). Proposed electrochemical biosensor displayed low detection limit (2.4 fM).

Keywords: CYFRA21-1; Electrochemical biosensor; Guanine oxidation; Label-free; Lung cancer.