In this study, we constructed a disposable indium tin oxide polyethylene terephthalate film (ITO-PET)-based electrochemical aptasensor for analyzing prostate-specific antigen (PSA), one of the most important biomarkers of prostate cancer. Because of their clinical importance, building PSA biosensing systems with high sensitivity and stability is essential. However, it still presents significant difficulties, such as low detection limits. We designed a platform to covalently bind the amino-terminated aptamer by modifying the ITO-PET surface with carboxyethylsilanetriol (CTES) to obtain a self-assembled monolayer (SAM). We also evaluated the potential for use in real human serum samples by investigating the optimal operating conditions and analytical performance characteristics of the developed biosensor. The design we present here exhibits excellent precision, with a limit of detection (LOD) as low as 8.74 fg/mL PSA. The broad linear detection range of the biosensor under optimal conditions was determined as 1.0-1500 fg/mL. The dissociation constant (Kd) for the aptamer was also calculated as 46.28 ± 5.63 nM by evaluating the impedimetric response as a function of PSA concentration. The aptasensor displayed considerable repeatability (1.3% RSD) and reproducibility (7.51% RSD) and good storage stability (98.34% of the initial activity for 8 weeks). Additionally, we demonstrated that the technique we developed was quite efficient in estimating the kinetics of aptamer-analyte interactions by determining the Kd and single-frequency impedance (SFI) data. In conclusion, we proposed a selective and sensitive biosensor with the potential for clinical application and superior performance in real serum samples.
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