Three-dimensional (3D) porous laser-scribed graphene (LSG) is a potential electrode material for flexible electrochemical sensors due to its high efficiency and low cost. However, the growth of nanoparticles usually depends on reducers or harsh conditions, which limit the application of flexible biosensors. To resolve this issue, 2D MXene nanosheets were applied to functionalize 3D LSG sheets with a C-O-Ti covalent crosslink and obtain an LSG-MXene hybrid scaffold. The shape-controlled Au-Pd bimetallic nanoparticles were synthesized on the surface of LSG-MXene by a self-reduction process at room temperature for 6 min to enhance its catalytic performance. As a proof of concept, the obtained hybrid nanocomposite was used to assess ascorbic acid (AA, 10–1600 μM), dopamine (DA, 12–240 μM), and uric acid (UA, 8–100, 200–800 μM). The fabricated sensor exhibited a low detection limit (S/N = 3) of 3 μM AA, 0.13 μM DA, and 1.47 μM UA. Furthermore, the Au-Pd/MXene/LSG was successfully applied to determine AA, DA, and UA in urine samples. Predictably, these outstanding performance sensors have great potential in flexible and wearable electronics.