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Gamma‐interferon production and quality in stoichiometric fed‐batch cultures of Chinese hamster ovary (CHO) cells under serum‐free conditions
Author(s) -
Xie Liangzhi,
Nyberg Gregg,
Gu Xuejun,
Li Hanyuan,
Möllborn Fredrik,
Wang Daniel I. C.
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(19971205)56:5<577::aid-bit11>3.0.co;2-9
Subject(s) - chinese hamster ovary cell , glycosylation , cell culture , fed batch culture , laboratory flask , biology , hamster , stoichiometry , biochemistry , interferon gamma , chemistry , in vitro , microbiology and biotechnology , fermentation , genetics , organic chemistry
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.