Dremel B A, Li S Y, Schmid R D
Department of Enzyme Technology, GBF (Gesellschaft für Biotechnologische Forschung GmbH), Braunschweig, Germany.
Biosens Bioelectron. 1992;7(2):133-9. doi: 10.1016/0956-5663(92)90018-i.
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).
本文描述了一种基于光纤检测氧消耗的流动注射分析（FIA）系统，该系统使用固定化葡萄糖氧化酶（GOD）和乳酸氧化酶（LOD）对动物细胞培养物中的葡萄糖和乳酸浓度进行在线监测。通过指示剂荧光的分子氧动态猝灭来测定氧的消耗。GOD和LOD固定在酶反应器中的可控孔径玻璃（CPG）上，酶反应器直接连接到覆盖氧电极的特殊设计的光纤流通池。该系统对0 - 30 mM葡萄糖呈线性，在30 mM时相对标准偏差（r.s.d.）为5%（五次测量）；对0 - 30 mM乳酸也呈线性，在30 mM时r.s.d.为5%（五次测量）。所使用的酶反应器在连续运行中稳定超过4周，每小时可分析多达20个样品。该系统已成功应用于用于生产重组人抗凝血酶III（AT - III）的动物细胞培养物中葡萄糖和乳酸浓度的在线监测。将FIA系统获得的在线测量结果与Yellow Springs Instrument Company（YSI）的葡萄糖和乳酸分析仪获得的离线结果进行了比较。
