Gamma-interferon production and quality in stoichiometric fed-batch cultures of Chinese hamster ovary (CHO) cells under serum-free conditions
✍ Scribed by Liangzhi Xie; Gregg Nyberg; Xuejun Gu; Hanyuan Li; Fredrik Möllborn; Daniel I. C. Wang
- Publisher
- John Wiley and Sons
- Year
- 1997
- Tongue
- English
- Weight
- 90 KB
- Volume
- 56
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
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, byproduct reduction, and Gamma-Interferon (IFN-␥) 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-␥ was monitored off-line during the culture process. The accumulative IFN-␥ 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-␥ glycosylation efficiency.