Sol–gel based glucose biosensors employing optical oxygen transducers, and a method for compensating for variable oxygen background

Abstract

Various types of thin-film glucose biosensors based on the use of the enzyme glucose oxidase (GOx) have been developed. The luminescent oxygen probe Ru(dpp) — whose emission is quenched by oxygen — is used to measure the consumption of oxygen. Three different combinations of oxygen transducer and sol–gel immobilized GOx were tested. In the first, GOx was sandwiched between a sol–gel layer doped with Ru(dpp) and a second sol–gel layer composed of pure sol–gel (the ‘sandwich’ configuration). In the second, a sol–gel layer doped with Ru(dpp) was covered with sol–gel entrapped GOx (the ‘two-layer configuration’). In the third, both GOx and a sol–gel powder containing GOx were incorporated into a single sol–gel phase (the ‘powder configuration’). In all cases, it was found to be essential to add sorbitol which results in a more porous sol–gel in which diffusion is not impaired. The sandwich configuration provides the highest enzyme activity and the largest dynamic range (0.1–15 mM), but suffers from a distinct decrease in sensitivity upon prolonged use. The two-layer configuration has the fastest response time ( t 90=50 s), while the ‘powder configuration’ provides the best operational lifetime. The storage stability of all configurations exceeds 4 months if stored at 4°C. In an Appendix, equations are derived which describe the response of such sensors, how the effect of varying oxygen supply can be compensated for by making use of two sensors, one sensitive to oxygen only, the other to both oxygen and glucose, and how such sensors can be calibrated using two calibrators only.