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软件FIACRE:基于流动注射分析,同时使用尿素光极和葡萄糖发光传感器进行生物过程监测。

Software FIACRE: bioprocess monitoring on the basis of flow injection analysis using simultaneously a urea optode and a glucose luminescence sensor.

作者信息

Busch M, Höbel W, Polster J

机构信息

Lehrstuhl für Allgemeine Chemie und Biochemie, Technischen Universität München, Freising-Weihenstephan, Germany.

出版信息

J Biotechnol. 1993 Dec;31(3):327-43. doi: 10.1016/0168-1656(93)90078-2.

Abstract

Various computer programs for large-scale bioprocess control and optimization have been developed as well as software for simple laboratory routine analysis. In comparison, software can hardly be found that works on laboratory scale and provides the control of complex flow injection analysis (FIA) systems, multisubstrate determination, data evaluation as well as minimal process control abilities. The sensors applied can be of different type (luminometric or other optical as well as electrochemical biosensors). The development of such a software may be very helpful for the transfer of FIA/biosensor systems from the state of development to industrial processes. Hence, each analysing system--even a well established biosensor--has to be individually adapted to the process, a task which is best done under laboratory conditions. Such a flexible, computer-controlled FIA system for research level based on the software FIACRE is presented. Five FIA/(bio)sensor system can be controlled simultaneously. Additionally, common temperature and pH recordings are possible. Determinations of substrate concentrations are performed by means of calibration curves which can be recorded at different times. This allows supervising the activities of the sensors during a cell cultivation and controlling the bioprocess, e.g. by adding substrate to a cell culture. The automated monitoring of the degradation of glucose and urea by two different optical sensing principles during a cell cultivation under the control of one microcomputer is presented for the first time. For this purpose, already well examined biosensors (a urease optode and a luminometric glucose sensor) were employed and their properties discussed under the aspect of working in real cultivation media. It will also be shown that substrates being of interest for bioprocess control can be detected by slight modifications of known reactions. For example, substrates of NADH-dependent enzymatic reactions can be detected by the luminol chemiluminescence system, and optodes can be employed for pH, penicillin and glucose determination.

摘要

已经开发出各种用于大规模生物过程控制和优化的计算机程序以及用于简单实验室常规分析的软件。相比之下,很难找到适用于实验室规模且能控制复杂流动注射分析(FIA)系统、多底物测定、数据评估以及具备最低限度过程控制能力的软件。所应用的传感器可以是不同类型的(发光或其他光学以及电化学生物传感器)。开发这样的软件对于将FIA/生物传感器系统从研发状态转移到工业过程可能会非常有帮助。因此,每个分析系统——即使是成熟的生物传感器——都必须针对该过程进行单独调整,这项任务在实验室条件下完成最为合适。本文介绍了一种基于软件FIACRE的用于研究水平的灵活的、计算机控制的FIA系统。可以同时控制五个FIA/(生物)传感器系统。此外,还可以进行常规的温度和pH记录。通过在不同时间记录的校准曲线来测定底物浓度。这使得在细胞培养过程中能够监测传感器的活性并控制生物过程,例如通过向细胞培养物中添加底物。首次展示了在一台微型计算机的控制下,在细胞培养过程中通过两种不同的光学传感原理对葡萄糖和尿素降解的自动监测。为此,使用了已经经过充分检验的生物传感器(一种脲酶光极和一种发光葡萄糖传感器),并在实际培养介质中工作的方面讨论了它们的特性。还将表明,通过对已知反应进行轻微修改,可以检测出对生物过程控制有意义的底物。例如,依赖NADH的酶促反应的底物可以通过鲁米诺化学发光系统进行检测,并且光极可用于pH、青霉素和葡萄糖的测定。

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