ABSTRACT
This paper described an impedimetric immunosensor for detecting Plasmodium falciparum histidine-rich protein 2 (PfHRP2). Antibodies from egg yolk (Ab-PfHRP2, IgY type) were linked covalently to the screen-printed gold electrodes (SPGE) surface modified with a thin film of Poly-pyrrole-pyrrole 3 carboxylic acid (P(Py-Py3COOH) to develop the sensing platform. The fabrication steps were followed by microscopic (scanning electron microscopy), spectroscopic (RAMAN spectroscopy and Energy-dispersive X-ray spectroscopy), and electrochemical (electrochemical impedance spectroscopy (EIS) and cyclic voltammetry) techniques. The determination of Ag-PfHRP2 was performed by EIS, and the BSA(bovine serum albumin)/Ab-PfHRP2(IgY)/P(Py-Py3COOH)/SPGE immunosensor recorded a linear response at 100-1000 ng mL-1 concentration range, with a limit of detection (LOD) of 27.47 ng mL-1. Its performance was confirmed by Enzyme-Linked Immuno-Sorbent Assay. The fabricated device uses a simple strategy of IgY immobilization, showing high sensitivity and good selectivity, and can be considered an alternative for carrying out malaria tests.
Subject(s)
Biosensing Techniques , Polymers , Biosensing Techniques/methods , Carboxylic Acids , Egg Yolk , Electrochemical Techniques/methods , Electrodes , Gold/chemistry , Histidine , Immunoassay/methods , Immunoglobulins , Limit of Detection , Polymers/chemistry , Pyrroles/chemistry , Serum Albumin, BovineABSTRACT
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.
