Premium
Multichannel flow‐injection‐analysis biosensor system for on‐line monitoring of glucose, lactate, glutamine, glutamate and ammonia in animal cell culture
Author(s) -
Blankenstein G.,
Spohn U.,
Preuschoff F.,
Thommes J.,
MR Kula
Publication year - 1994
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1994.tb00319.x
Subject(s) - biosensor , glutamine , chemiluminescence , flow injection analysis , glucose oxidase , chemistry , chromatography , cell culture , luminol , biochemistry , enzyme , bioprocess , immobilized enzyme , biology , amino acid , detection limit , paleontology , genetics
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.