An automatic dehydrogenase-based flow-injection system: application for the continuous determination of glucose and lactate in mammalian cell-cultures.
A concept for the development of an automatic flow-injection analyzer with integrated dehydrogenase columns and its application in the control of industrial processes is presented. The system is based upon a kernel consisting of a nested-loop injection unit, pumps for the filling of the injection loops and the transport of buffer and values for switching on the one hand between sample and standard solutions and on the other hand between different enzyme columns. A Microsoft Windows 3.x application 'WIN-FIA' controls interactively the whole system and can be easily adapted to a specific solution of an analytical problem. As an example, the flow-injection system was used for the continuous determination of glucose and lactate, using glucose dehydrogenase (GDH) and lactate dehydrogenase (LDH) as indicator enzymes, in a mammalian cell-culture fermentation process. The resulting concentration values are in good agreement with those obtained by discontinuously taken standard spectrophotometric enzyme assays.
Becker
T
T
Lehrstuhl für Allgemeine Chemie und Biochemie, Technische Universität München, Freising-Weihenstephan, Germany.
Schuhmann
W
W
Betken
R
R
Schmidt
H L
HL
Leible
M
M
Albrecht
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D004800:Enzymes, Immobilized; D005285:Fermentation; D017022:Flow Injection Analysis / Q000295:instrumentation; D005947:Glucose / Q000032:analysis*; D042843:Glucose 1-Dehydrogenase; D005948:Glucose Dehydrogenases / Q000032:analysis*; D006825:Hybridomas / Q000737:chemistry*; D007202:Indicators and Reagents; D007770:L-Lactate Dehydrogenase / Q000032:analysis*; D007773:Lactates / Q000032:analysis*; D012984:Software
D000911:Antibodies, Monoclonal; D004800:Enzymes, Immobilized; D007202:Indicators and Reagents; D007773:Lactates; D005948:Glucose Dehydrogenases; D007770:L-Lactate Dehydrogenase; D042843:Glucose 1-Dehydrogenase; D005947:Glucose