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The design, fabrication, and in-vitro evaluation of an amperometric biochip that is designed for the continuous in vivo monitoring of physiological analytes is described. The 2 ×4 ×0.5 mm biochip contains two platinum working enzyme electrodes that adopt the microdisc array design to minimize diffusional limitations associated with enzyme kinetics. This configuration permits either dual analyte sensing or a differential response analytical methodology during amperometric detection of a single analyte. The working enzyme electrodes are complemented by a large area platinized platinum counter electrode and a silver reference electrode. The biorecognition layer of the working electrodes was fabricated from around 1.0-μm-thick composite membrane of principally tetraethylene glycol (TEGDA) cross-linked poly(2-hydroxyethyl methacrylate) that also contained a derivatized polypyrrole component and a biomimetic methacrylate component with pendant phosphorylcholine groups. These two additional components were introduced to provide interference screening and in vivo biocompatibility, respectively. This composite membrane was used to immobilize glucose oxidase and lactate oxidase onto both planar and microdisc array electrode designs, which were then used to assay for in vitro glucose and lactate, respectively. The glucose biosensor exhibited a dynamic linear range of 0.10-13.0 mM glucose with a response time (t95) of 50 s. The immobilized glucose oxidase within the hydrogel yielded a Km(app) of 35 mM, not significantly different from that for the native, solution-borne enzyme (33 mM). The microdisc array biosensor displayed linearity for assayed lactate up to 90 mM, which represented a 30-fold increase in linear dynamic lactate range compared to the biosensor with the planar electrode configuration. Preliminary in vitro operational stability tests performed with the microdisc array lactate biosensor demonstrated retention of 80% initial biosensor response after five days of continuous operation in buffer under physiologic conditions of pH and temperature.