Deposition of glucose oxidase on the surface of microtiter plate: its application to determination of glucose in foods

Abstract

A facile method for the immobilization of enzyme on the surface of the 96-well microtiter plate has been studied using avidin and biotinylated glucose oxidase (GOD). It was demonstrated that an alternate and repeated deposition of avidin and biotinylated GOD gave an activity dependent on the number of GOD layers deposited. The well, after deposition of 16 layers of GOD, was capable of determining glucose up to 1 mM. The enzyme activity of the well was retained for 6 months when the plate was kept at room tempearature. No significant decrease in the activity was observed for 50 successive determinations of 1 mM glucose. These results suggest that the avidin/biotin system would be feasible to the immobilization of enzymes on the surface of a microtiter plate. The determination of glucose in various food samples using the above method is also described.