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The development of nano biocomposite film (f-MWCNTs-Au-GOx) for L-cysteine (LC) detection is?proposed byusing glassycarbon electrode (GCE). The proposed nano biocomposite film has been?fabricated on ITO for scanning electron microscopy(SEM) and atomic force microscopy(AFM) analysis.?Next, the f-MWCNTs-Au-GOx nano biocomposite film modified GCE’s surface was examined?byscanning electrochemical microscopy(SECM). The proposed nano biocomposite film has been?successfullyapplied for the detection of LC using cyclic voltammetry(CV) and amperometry. Thef-MWCNTs-Au-GOx film modified GCE exhibited a linear response for LC detection in the lower?and higher concentrations ranges of 2 to 42×10?6, 0.1 to 1.08×10?3 mol L?1. Also, the proposed?nano biocomposite film possesses high sensitivityand good repeatabilityf or LC detection.