On-line determination of glucose and lactate concentrations in animal cell culture based on fibre optic detection of oxygen in flow-injection analysis.
A flow-injection analysis (FIA) system based on fibre optic detection of oxygen consumption using immobilized glucose oxidase (GOD) and lactate oxidase (LOD) is described for the on-line monitoring of glucose and lactate concentrations in animal cell cultures. The consumption of oxygen was determined via dynamic quenching by molecular oxygen of the fluorescence of an indicator. GOD and LOD were immobilized on controlled pore glass (CPG) in enzyme reactors which were directly linked to a specially designed fibre optic flow-through cell covering the oxygen optrode. The system is linear for 0-30 mM glucose, with an r.s.d. of 5% at 30 mM (five measurements) and for 0-30 mM lactate, with an r.s.d. of 5% at 30 mM (five measurements). The enzyme reactors used were stable for more than 4 weeks in continuous operation, and it was possible to analyse up to 20 samples per hour. The system has been successfully applied to the on-line monitoring of glucose and lactate concentrations of an animal cell culture designed for the production of recombinant human antithrombine III (AT-III). Results of the on-line measurement obtained by the FIA system were compared with the off-line results obtained by a glucose and lactate analyser from Yellow Springs Instrument Company (YSI).
Dremel
B A
BA
Department of Enzyme Technology, GBF (Gesellschaft für Biotechnologische Forschung GmbH), Braunschweig, Germany.
Li
S Y
SY
Schmid
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RD
D000818:Animals; D015374:Biosensing Techniques; D002138:Calibration; D002478:Cells, Cultured; D004800:Enzymes, Immobilized; D005285:Fermentation / Q000502:physiology; D005336:Fiber Optic Technology; D017022:Flow Injection Analysis; D005947:Glucose / Q000032:analysis*; D007773:Lactates / Q000032:analysis*; D019344:Lactic Acid; D010100:Oxygen / Q000032:analysis*; D013050:Spectrometry, Fluorescence
D004800:Enzymes, Immobilized; D007773:Lactates; D019344:Lactic Acid; D005947:Glucose; D010100:Oxygen