Polyimide-laser-engraved porous graphene (LEPG) are hopeful electrode modification materials for flexible electrochemical sensing based on its high-efficiency preparation and low cost. Herein, a flexible, multi-patterned, and miniaturized electrode was fabricated via a simple and novel direct laser engraving. 3D LEPG with porous network structure can selective decorated with Pt nanoparticles (Pt NPs) by in situ electrochemical depositions (Pt-LEPG) as sensitively HO sensors with a wide range of linear (0.01-29 nM) and high sensitivity (575.75 μA mM cm). Subsequently, a glucose biosensor was successfully constructed through immobilized glucose oxidases (GOD) onto Pt-LEPG electrode. New-designed GOD/Pt-LEPG glucose sensor exhibited a noteworthy lower limit of detection (0.3 μM, S/N = 3) and high sensitivity (241.82 μA mM cm), as much a wide-range of linear (0.01-31.5 mM) at near-neutral pH conditions, enabling detect glucose in real human serum specimens with satisfactory results. Predictably, these outstanding performance sensors have great potential in terms of flexible and wearable electronics.