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Cost-effective valorization of biomass into advanced carbon remains a challenge. Here we reported a facile and ultrafast laser writing technique to convert biomass into porous graphene for electrochemical sensing. Laser-induced graphene (LIG) was synthesized from a fully biomass-based film composed of kraft lignin (KL) and cellulose nanofibers (CNFs). The LIG-based electrode was applied to detect dopamine using cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques. Dopamine with a concentration ranging from 5 to 40 μM was detected linearly, with a sensitivity of 4.39 μA μM −1 cm −2 . Our study eliminated the use of synthetic polymer for lignin-based film formation. It demonstrated the feasibility of using the film fully composed of biomass for LIG formation. Furthermore, derived LIG electrodes were shown to have high electrochemical sensing performance.