Huang Y L, Li S Y, Dremel B A, Bilitewski U, Schmid R D
Department of Enzyme Technology, Gesellschaft für Biotechnologische Forschung GmbH, Braunschweig, F.R.G.
J Biotechnol. 1991 Apr;18(1-2):161-72. doi: 10.1016/0168-1656(91)90244-p.
A flow-injection analysis (FIA) system for the on-line determination of glucose in animal cell cultures is described. The system is based on immobilized glucose oxidase (GOD). The hydrogen peroxide generated in the enzyme reaction is determined via a highly sensitive chemiluminescent reaction with luminol. Based on the measurement of the maximum emitted light intensity, the system was able to analyse hydrogen peroxide over the concentration range of 10(-7) to 10(-2) M. For glucose determination, the system has a linear range of 10(-5) to 5 x 10(-2) M glucose, with an r.s.d. of 3% at the 1 mM level (5 measurements). The influence of luminol and buffer concentrations, pH and temperature on the chemiluminescent reaction were investigated. The enzyme reactor used was stable for more than 4 weeks in continuous operation, and it was possible to analyse up to 20 samples per h. The system has been successfully applied to on-line monitoring of glucose concentration during an animal cell culture, designed for the production of human antithrombin III factor. Results obtained with the FIA system were compared with off-line results, obtained with a Yellow Springs Instrument Company Model 27 (YSI).
本文描述了一种用于在线测定动物细胞培养液中葡萄糖的流动注射分析（FIA）系统。该系统基于固定化葡萄糖氧化酶（GOD）。酶反应中产生的过氧化氢通过与鲁米诺的高灵敏度化学发光反应进行测定。基于对最大发射光强度的测量，该系统能够分析浓度范围为10^(-7)至10^(-2) M的过氧化氢。对于葡萄糖测定，该系统的线性范围为10^(-5)至5×10^(-2) M葡萄糖，在1 mM水平（5次测量）时相对标准偏差为3%。研究了鲁米诺和缓冲液浓度、pH值和温度对化学发光反应的影响。所使用的酶反应器在连续运行中稳定超过4周，每小时最多可分析20个样品。该系统已成功应用于为生产人抗凝血酶III因子而设计的动物细胞培养过程中葡萄糖浓度的在线监测。将FIA系统获得的结果与使用Yellow Springs Instrument Company Model 27（YSI）获得的离线结果进行了比较。
