Optical fibre biosensors using enzymatic transducers to monitor glucose

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Abstract

The construction and performance of a novel enzyme based optical sensor for in situ continuous monitoring of glucose in biotechnological production processes is presented. Sensitive optical coatings are formed from inorganic?organic hybrid polymers (ORMOCER®s*) combined with a flurophore (ruthenium complex) and an enzyme, and applied to lenses, declad polymer optical fibre (POF) and polymer clad silica fibre (PCS). The enzyme, glucose oxidase, catalyzes oxidization of glucose to gluconic acid by depleting oxygen. Oxygen consumption is determined by measuring the fluorescence lifetime of metal organic ruthenium complexes which are quenched by oxygen. The coatings developed were designed to adhere to glass and polymer surfaces, to be compatible with enzymes and ruthenium complexes, and were demonstrated both as double- and single-layer structures. The sensor response to gaseous oxygen, dissolved oxygen and dissolved glucose was measured via fluorescence lifetime changes. A best detection limit of 0.5% (vol) has been determined for gaseous O2 with selected ORMOCER® sensing layers. Glucose concentrations were measured to a detection limit of 0.1 mmol L−1 over a range up to 30 mmol L−1. The sensor was usable for 30 days in a bioreactor. The opto-electronic instrumentation and performance in laboratory bioreactors and in an industrial reactor are evaluated.

Keyword(s)

optical fibre sensor, hybrid coating, biosensor, enzyme, glucose, oxygen, glucose oxidase, polymer, POF, bioreactor, ruthenium, fluorescence, fluorophore

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