In this paper, we present a neutral non-enzymatic glucose biofuel cell (GBFC) based on a Pt/Au nano-alloy anode. We synthesize Pt/Au alloy anodes by the electrochemical deposition of a Pt/Au nano-alloy on the surface of a thin polyethylene terephthalate membrane with a uniformly distributed microhemispheric array. The experimental results demonstrate that the proposed GBFC can generate a maximum power density of 0.32 mW/cm2, an open-circuit voltage (Voc) of 0.42 V, and a short-circuit current (Isc) density of 2.67 mA/cm2 with 5 mM of glucose in a 0.1-M PBS (pH 7.4) solution while the cathode is being supplied with 7% saturated oxygen. The synergistic effect of the Pt/Au bimetallic alloy is found to improve the performance of the proposed GBFC. The proposed neutral non-enzymatic GBFC shows promising feasibility as an implantable power supply system for the provision of continuous and consistent electricity to a continuous glucose monitoring system.