In this paper, a simple method for immobilization of acetylcholinesterase (AChE) on cysteamine assembled glassy carbon electrode coupled with gold nanoparticles (GNPs) was proposed and thus a sensitive, fast and stable amperometric biosensor for quantitative determination of monocrotophos was developed. The fabrication procedure was characterized by cyclic voltammetry, electrochemical impedance spectroscopy and contact angles. The presence of GNPs not only led to an increased effective surface to provide a sufficient amount of sites for binding enzyme, but also promoted electron transfer react...