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Implantable artificial kidneys represent a promising alternative for patients with end-stage renal disease (ESRD), aiming to overcome the limitations of conventional dialysis through the integration of microfluidic and electrokinetic technologies. In this study, we present a sawtooth electrode microfluidic chamber that achieves blood cell separation via negative dielectrophoresis at a record-low operating voltage of 1.4 V, representing a fivefold reduction compared with rectangular electrode designs and supporting potential integration into implantable artificial kidney systems. A microfluidic chip incorporating an asymmetric sawtooth electrode geometry was developed to enhance local electric field gradients while reducing power consumption. Device performance was investigated using COMSOL Multiphysics simulations. Response Surface Methodology (RSM) based on a Box–Behnken design was employed to optimize the number of teeth per unit length (N), sawtooth height (H), and applied voltage (V), while excitation frequency was fixed at 1 MHz and flow velocity was maintained constant at 0.1 µL·min−1. Statistical analysis was conducted using analysis of variance (ANOVA) in Minitab (Version 27; Minitab, LLC, State College, PA, USA, 2024). The optimization model showed strong predictive capability (R2 = 95.8%) and identified applied voltage (59.45% contribution) and sawtooth height (33%) as the dominant factors affecting separation efficiency, with a significant H × V interaction (p = 0.023). Comprehensive voltage-response mapping over the range of 0.8–4.0 V revealed four operational regimes, including a previously unreported high-voltage failure zone above 2.8 V, where electrothermal flow and electroporation degrade performance. Under physiological conductivity conditions, the optimized design maintained a separation efficiency of 78.3% at 1.4 V with a tip temperature rise of only 1.2 °C, while full recovery of performance was achieved at 2.2 V. Cell-specific separation efficiencies reached 97.3% for white blood cells, 95.8% for red blood cells, and 84.7% for platelets, reducing the downstream cellular load by 92.6%. These findings demonstrate that the proposed low-voltage, high-efficiency separation platform has strong potential as a cellular pre-filtration module in implantable artificial kidney systems and other lab-on-chip biomedical devices.