In the present work, we describe a novel one-step enzyme-free dual electrochemical immunosensor for the determination of histidine-rich protein 2 (Ag-PfHRP2), a specific malaria biomarker. A gold electrode (GE) was functionalized with the PfHRP2 antibody (Ab-PfHRP2) using dihexadecyl phosphate (DHP) polymer as an immobilization platform. The Ab-PfHRP2/DHP/GE sensor was characterized by cyclic voltammetry, electrochemical impedance spectroscopy, Fourier-transform infrared spectroscopy, scanning electron microscopy, and atomic force microscopy. The developed immunosensor was employed for indirect Ag-PfHRP2 determination by differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). The linear range was 10-400 ng mL-1 and 10-500 ng mL-1 for EIS and DPV, while the limit of detection was 3.3 ng mL-1 and 2.8 ng mL-1, respectively. The electrochemical immunosensor was successfully applied for Ag-PfHRP2 determination in human serum samples. Its performance was compared with an ELISA test, and good correspondence was achieved. The coefficients of intra- and inter-assay variations were less than 5%. The electrochemical immunosensor is a useful and straightforward tool for in situ malaria biomarker determination.
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