In this research, a novel interdigitated gear-shaped, graphene-based electrochemical biosensor was developed for the detection of dopamine (DA). The sensor’s innovative design improves the active surface area by 94.52% and 57% compared to commercially available Metrohm DropSens 110 screen-printed sensors and printed circular sensors, respectively. The screen-printed electrode was fabricated using laser processing and modified with graphene polyaniline conductive ink (G-PANI) to enhance its electrochemical properties. Fourier Transform Infrared (FTIR) Spectroscopy and X-ray diffraction (XRD) were employed to characterize the physiochemical properties of the sensor. Dopamine, a neurotransmitter crucial for several body functions, was detected within a linear range of 0.1–100 µM, with a Limit of Detection (LOD) of 0.043 µM (coefficient of determination, R² = 0.98) in phosphate-buffer saline (PBS) with ferri/ferrocyanide as the redox probe. The performance of the sensor was evaluated using cyclic voltammetry (CV) and Chronoamperometry, demonstrating high sensitivity and selectivity. The interdigitated gear-shaped design exhibited excellent repeatability, with a relative standard deviation (RSD) of 1.2% (n = 4) and reproducibility, with an RSD of 2.3% (n = 4). In addition to detecting dopamine in human serum, the sensor effectively distinguished dopamine in a ternary mixture containing uric acid (UA) and ascorbic acid (AA). Overall, this novel sensor design offers a reliable, disposable, and cost-effective solution for dopamine detection, with potential applications in medical diagnostics and neurological research.