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On‐line monitoring of glucose in mammalian cell culture using a flow injection analysis (FIA) mediated biosensor
Author(s) -
Male Keith B.,
Gartu P. Octavian,
Kamen A. Amine,
Luong John H. T.
Publication year - 1997
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19970805)55:3<497::aid-bit5>3.0.co;2-7
Subject(s) - biosensor , glucose oxidase , chromatography , flow injection analysis , bioreactor , amperometry , chemistry , detection limit , immobilized enzyme , hexokinase , biochemistry , electrode , enzyme , organic chemistry , glycolysis , electrochemistry
A flow injection analysis (FIA) biosensor system has been developed for on‐line determination of glucose during mammalian cell cultivation. The culture sample was peristaltically withdrawn from the bioreactor and after cell separation by a steam sterilizable ceramic microfilter, the filtrate was continuously fed to the FIA mediated‐biosensor system at 4 mLh −1 , whereas the cell‐containing retentate was recirculated to the bioreactor. In the amperometric biosensor system, glucose oxidase was covalently immobilized onto a preactivated nylon membrane and attached to the sensing area of a platinum working electrode. The enzyme reaction was coupled with the mediator 1,1′‐dimethylferricinium (DMFe + )‐cyclodextrin inclusion complex to recycle the reduced glucose oxidase to its original active state. 1,1′‐Dimethylferrocene (DMFe) was then reoxidized to DMFe + at the surface of the platinum electrode poised at + 0.15 V vs silver/silver chloride. The FIA mediated‐biosensor was linear up to 6 mM glucose, with a detection limit of 0.1 mM, and possessed excellent reproducibility (± 0.4 %, 95 % confidence interval) over 123 repeated analyses during a 62 h continuous operation. The immobilized glucose oxidase was stable for up to 7 days when applied to glucose measurement during 5–10 day fed‐batch cultivation of 293S mammalian cells. The results obtained from the mediated‐biosensor system compared well with the hexokinase and HPLC data. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55 : 497–504, 1997