Multichannel flow-injection-analysis biosensor system for on-line monitoring of glucose, lactate, glutamine, glutamate and ammonia in animal cell culture.
The application of a chemiluminometric method for the on-line monitoring of a hybridoma cell culture is described. Enzyme sensors for glucose, lactate, glutamine, glutamate and ammonia, based on oxidase-catalysed reactions, were developed and connected to a flow-injection-analysis (f.i.a.) biosensor. H2O2 produced by the oxidase-catalysed enzyme reaction was detected by luminol chemiluminescence with a fibre-optic H2O2 biosensor. The system has been used to monitor animal cell cultures. A continuous hybridoma cell cultivation for the production of monoclonal antibodies is presented as an example. It was possible to monitor the bioprocess over a period of 15 days. A complete analysis of all five components could be performed within 42 min. The enzyme sensors were stable during the whole cultivation time without significant loss of activity. The computer-controlled biosensor f.i.a. works with good reliability. The precision for all five components ranged between 2.2 and 4.5%. It was possible to determine glutamine in one step using an anti-interference enzyme reactor. Endogenous glutamate was completely removed up to a level of 0.5 mM.
Blankenstein
G
G
Institute of Enzyme Technology, Heinrich-Heine University Düsseldorf, Jülich, Germany.
Spohn
U
U
Preuschoff
F
F
Thömmes
J
J
Kula
M R
MR
D000641:Ammonia / Q000032:analysis*; D000818:Animals; D015374:Biosensing Techniques; D002478:Cells, Cultured; D017022:Flow Injection Analysis; D005947:Glucose / Q000032:analysis*; D018698:Glutamic Acid / Q000032:analysis*; D005973:Glutamine / Q000032:analysis*; D006825:Hybridomas / Q000378:metabolism; D007773:Lactates / Q000032:analysis*; D019344:Lactic Acid; D008163:Luminescent Measurements; D051379:Mice; D009862:Online Systems
D007773:Lactates; D005973:Glutamine; D019344:Lactic Acid; D018698:Glutamic Acid; D000641:Ammonia; D005947:Glucose