In this study, we developed an electrochemical-based single-use neurobiosensor based on multiwalled carbon nanotube (MWCNT)-gold nanoparticle (AuNP) nanocomposite doped, 11-amino-1-undecanethiol (11-AUT)-modified polyethylene terephthalate coated indium tin oxide (ITO-PET) electrodes. This electrode was used for the sensitive determination of DJ-1, a protein responsible for mitochondrial dysfunction in Parkinson’s disease (PD) with the task of eliminating oxidative stress. The design strategy and analytical studies for the neurobiosensor were monitored with electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and single frequency impedance (SFI) techniques. The selective determination range for DJ-1 of the developed neurobiosensor system is 4.7-4700 fg mL in accordance with the charge transfer resistance (Rct) associated with a limit of detection of 0.5 fg mL. Since changes in the expression of DJ-1 protein is particularly important in cerebrospinal fluid (CSF) and saliva, the ability of the developed neurobiosensor system to detect the DJ-1 protein in these media was tested by the standard addition method. The statistical results show that the biosensor decorated with MWCNT-AuNP-AUT may be recommended for the selective determination of DJ-1 protein.Copyright © 2020 Elsevier B.V. All rights reserved.