Label-free electrochemical aptasensor for progesterone detection in biological fluids

A label-free electrochemical progesterone (P4) aptasensor was successfully developed by covalently immobilizing NH₂-functionalized P4-specific aptamer on the electrode surface. The NiO-Au hybrid nanofibers were synthesized by the electrospinning technique. GQDs-NiO-AuNFs nanocomposite was prepared by dispersing of electrospun NiO-AuNFs in the as-synthesized graphene quantum dots (GQDs) solution and stirring for 24 h. Novel GQDs-NiO-AuNFs nano-architecture in combination with functionalized multiwalled carbon nanotubes (f-MWCNTs) were further utilized to modify screen printed carbon electrode (SPCE) in order to construct an effective immobilization matrix with plenty of carboxylic functional groups. The stepwise assembly process of the designed aptasensor was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The aptamer-progesterone complex formation led to a hindered electron transfer reaction on the sensing interface, which decreased the redox probe peak current. Based on of this, progesterone could be quantitatively detected by monitoring the decrease of differential pulse voltammetric (DPV) responses of [Fe(CN)₆]^3-/4- peak current with increasing the progesterone concentration. Under optimized experimental parameters, the aptasensor exhibited a dynamic concentration range from 0.01 to 1000 nM and a detection limit of 1.86 pM. The proposed aptasensor was successfully employed for the determination of progesterone in human serum samples and pharmaceutical formulations.