AbstractsBiology & Animal Science

Luminescent microspheres encapsulating glucose oxidase and an oxygensensitive lumophore have recently been reported as potential implantable sensors for in vivo glucose monitoring. However, there are two main issues that must be addressed for enzymatic systems such as these to realize the goal of minimally-invasive glucose monitoring. The first issue is related to the short response range of such sensors, less than 200 mg/dL, which must be extended to cover the full physiological range (0-600 mg/dL) of glucose possible for diabetics. The second issue is concerning the short operating lifetime of these systems due to enzyme degradation (less than 7 days). Two approaches were considered for increasing the range of the sensor response; nanofilm coatings and particle porosity. In the first approach, microparticle sensors were coated with layer-by-layer deposited thin nanofilms to increase the response range. It was observed that, a precise control on the response range of such sensors can be achieved by manipulating different characteristics (e.g., thickness, deposition condition, and the outermost capping layer) of the nanofilms. However, even with 15 bilayers of poly(allylamine hydrochloride)/poly(styrene sulfonate) (PAH/PSS) nanofilm, limited range was achieved (less than 200 mg/dL). By performing extrapolation on the data obtained for the experimentally-determined response range versus the number of PAH/PSS bilayers, it was predicted that a nanofilm coating comprising of more than 60 PAH/PSS bilayers will be needed to achieve a linear response up to 600 mg/dL. Using modeling, it was realized that a more effective method for achieving a linear response up to 600 mg/dL is to employ microparticles with higher porosity. Sensors were prepared from highly porous silica microparticles (diameter = 7 mu m, porosity = 0.6) and their experimental response was determined. Not surprisingly, the experimentally determined response range of such sensors was found to be higher than 600 mg/dL. To improve the longevity of these sensors, two approaches were employed; incorporation of catalase and increasing the loading of glucose oxidase. Catalase was incorporated into microparticles, which protects the enzyme from peroxide-mediated deactivation, and thus improves the stability of such sensors. Sensors incorporating catalase were found to ~5 times more stable than the GOx-only sensors. It was theoretically predicted, that by maximizing the loading of glucose oxidase within the microparticles, the longevity of such sensors can be substantially improved. Based on this understanding, sensors were fabricated using highly porous microparticles; response range did not vary even after one month of continuous operation under normal physiological conditions. Modeling predicts that 1 mM of glucose oxidase and 1 mM of catalase would extend the operating lifetime to more than 90 days.