ISSN:
0006-3592
Keywords:
fed-batch
;
gamma-Interferon
;
CHO cells
;
glycosylation
;
glucose starvation
;
protein quality
;
nutritional control
;
Chemistry
;
Biochemistry and Biotechnology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Process Engineering, Biotechnology, Nutrition Technology
Notes:
The application of a stoichiometric medium design approach was studied in fed-batch cultivation of Chinese hamster ovary (CHO) cells. A serum-free medium containing a very low protein concentration (2 mg/L insulin) was developed. A supplemental medium was formulated according to the stoichiometric equation governing cell growth using cell composition obtained from hybridoma cells. Fed-batch culture was conducted in spinner flasks using the supplemental medium for feeding. Significant improvement in cell growth, by-product reduction, and Gamma-Interferon (IFN-γ) production was achieved as compared to a typical batch culture. Results indicate that the stoichiometric approach, originally developed for hybridoma cultures, is a fast and effective method for cell culture process design and improvement. The glycosylation of IFN-γ was monitored off-line during the culture process. The accumulative IFN-γ glycosylation efficiency was slightly improved as compared to that of the batch culture, due to the nutritional control through the stoichiometric feeding. Periodic glucose starvation was observed during the fed-batch culture as a result of the manual feeding. Pulse-chase radiolabeling assay shows that glucose starvation leads to a deteriorated IFN-γ glycosylation efficiency. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 577-582, 1997.
Additional Material:
4 Ill.
Type of Medium:
Electronic Resource
