Each electrode chip was fabricated by a semiconducting processes including a photoresist coating, patterning, lift off, and passivation. As can be seen in Fig. 1(a), approximately 250 chips on a 4 in glass wafer were fabricated for each process. A central circle shaped Au electrode with an area of π mm2 was utilized as the working electrode ( Fig 1(b)). An Au-electrode printed circuit board Cyclopamine mouse (PCB) chip was fabricated by an electroplating method and two types of PCB chips were made for use in other applications ( Fig 1(c and d)). Functionalization of GO nanosheets with MPHs was achieved by following a previous

study Veerapandian et al. Briefly, 200 μL of MPHs and 40 μL of 3-APTES were added to a tube containing anhydrous C2H5OH and kept for 10 h. After completion of the reaction process, FGO was drop-cast onto the oxygen plasma cleaned Au electrode PCB chip and allowed to evaporate at room temperature for 1 h. After modification of each Au electrode on the wafer, multiple layers were spin coated on the wafer. These layers were composed of a silane coupling layer on top of the FGO-Au electrode

followed by GOX composites, nafion, a silane coupling layer, and a restricted permeable polymer layer to form the multilayer-FGO-Au electrode. A customized reading platform was designed and built for the experiment. Fig. 2(a) shows the layout of the read out main board, the capable analog signal range of the system is between +5 and −5 V. As can be seen in Fig. 2(b), indicated with arrow, five different chips can be placed into the slots Inhibitor Library supplier for performing simultaneous measurements of different concentrations of glucose in TES and urine and between-run tests in same

concentration of glucose. All experiments were performed at room temperature in a 5 mL of collected urine samples and TES buffer for characterization. All amperometric measurements were performed at a working electrode potential of +0.6 V. The concentration of glucose in the TES buffer was examined by the fabricated Au-PCBs and the customized platform and the resultant images are displayed in Fig. 3. As can be seen in Fig. 3(a), the amperometric response with Niclosamide glucose concentration has strong proportionality to the concentration as it increases. Fig. 3(b) shows the amperometric responses measured 7 s after the immersion of five different chips with their variations from the mean values on each concentration. The between-run results show that their variations are within 6% from their mean values. Such materials including Ag/AgCl and Pt were not used as reference and auxiliary electrodes in this study despite these being the conventionally used materials in most electrochemical systems. Instead, we used an Au substrate as reference and auxiliary to reduce cost of chip production and increase stability during the field measurements.