ISSN:
0006-3592
Keywords:
glucose
;
glucose oxidase
;
biosensor
;
on-line monitoring
;
1,1′-dimethylferrocene
;
mammalian cells
;
Chemistry
;
Biochemistry and Biotechnology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Process Engineering, Biotechnology, Nutrition Technology
Notes:
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 The culture sample was peristaltically withdrawn from the bioreactor and after cell separation by a steam sterilizable ceramic microfilter, the filtrate was continuously fed to the FIA mediated-biosensor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
Additional Material:
7 Ill.
Type of Medium:
Electronic Resource
Permalink
