A novel non-enzymatic glucose sensor was developed based on sandpaper-supported nickel coatings (SNC). Superhydrophilic network was formed on the surface of sandpaper via a simple laser-scribing (LS) method. Orthogonal model was utilized to optimize the processing parameters for preparing SNC with an optimal conductivity of 4.20 x 10(6) S.m(-1). The surface morphology, functional groups and elements of SNC were characterized by SEM, ATR-FTIR and XPS respectively. The crystallinity of nickel nanocoating was determined by XRD. Remarkably, the SNC exhibited favorable electrocatalytic performance with a wide linear range (LR) of 0.01-4.3 mM, a high sensitivity of 1163.3 mu A mM(-1) cm(-2) and a low limit of detection (LOD) of 0.250 mu M, which was measured by standard electrochemical measurements. Low relative standard deviation (0.59% and 0.72%, n = 6) and high recoveries (96.30-97.78%) verified its stability and reusability, which can meet the needs of daily uses. Finally, successful application of the SNC to the analysis of glucose in human serum samples, leading to a promising tool for non-enzymatic glucose determination in healthcare and food industries.