A facile method of preparing water-dispersible sulfonated graphene (SPG) using sulfonated poly(ether-ether-ketone) organic polymer as a modifier was realized. A glucose biosensor was fabricated by immobilizing glucose oxidase (GO(x)) on the surface of AuNPs used to modify SPG and chitosan (CH) deposited on an indium tin-oxide (ITO) glass electrode by a solution casting method. Morphological and structural characterizations confirm that the AuNPs can be efficiently applied to the SPG-CH matrix. The amperometric response of the GO(x)/SPG-AuNPs-CH/ITO bioelectrode shows a broad linear range of 0.5 to 22.2 mM, with a limit of detection of 0.13 mM and a high sensitivity of 6.51 mu A/(mM cm(2)). The excellent performance of the constructed biosensor is attributed to the large surface-to-volume ratio and electron transfer ability of SPG, the high catalytic activity of the AuNPs, and the good biocompatibility of CH. In addition, the sensor has important advantages, such as its simple preparation, fast response time (10 s), good stability (70 days), and high reproducibility. Favorable results upon examining the electrochemical response for the determination of glucose in human blood serum were obtained, without the assistance of a negligible effect of interfering bio-analytes. The results of studies show that the ternary SPG-AuNPs-CH nanocomposite may offer a new approach for developing novel types of highly sensitive and stable electrochemical biosensors. (C) 2013 Elsevier Ltd. All rights reserved.