On-line monitoring of glucose in mammalian cell culture using a flow injection analysis (FIA) mediated biosensor
✍ Scribed by Keith B. Male; P. Octavian Gartu; A. Amine Kamen; John H. T. Luong
- Publisher
- John Wiley and Sons
- Year
- 1997
- Tongue
- English
- Weight
- 200 KB
- Volume
- 55
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
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 The culture sample was peristaltically withdrawn from the bioreactor and after cell separation by a steam sterilizable ceramic microfilter, the filtrate was continuously fed to the FIA mediated-biosensor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 cellcontaining 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 1,1Ј-dimethylferricinium (DMFe + )-cyclodextrin inclusion complex to recycle the reduced glucose oxidase to its original active state. 1,1Ј-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.