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Influence of Primatone RL supplementation on sialylation of recombinant human interferon‐γ produced by Chinese hamster ovary cell culture using serum‐free media
Author(s) -
Gu Xuejun,
Xie Liangzhi,
Harmon Bryan J.,
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(19971120)56:4<353::aid-bit1>3.0.co;2-n
Subject(s) - chinese hamster ovary cell , cell culture , hydrolysate , extracellular , glycosylation , biochemistry , recombinant dna , cell growth , biology , fed batch culture , chemistry , hydrolysis , fermentation , genetics , gene
Although serum‐free media have been widely used in mammalian cell culture for therapeutic protein production, the effects of serum‐substitutes on product quality have not been extensively examined. This study observed an adverse effect of Primatone RL, an animal tissue hydrolysate commonly used as a serum‐substitute to promote cell growth, on sialylation of interferon‐γ (IFN‐γ) derived from Chinese hamster ovary (CHO) cell culture in both batch and fed‐batch modes. In batch cultures, decreased sialylation was observed at each of the glycosylation sites ( i.e., Asn 25 and Asn 97 ) of IFN‐γ with the use of elevated concentrations of the peptone. Although poorest sialylation was obtained with the use of a growth‐inhibiting concentration of Primatone RL, diminished sialylation was observed at the optimal peptone concentration for cell growth and product yield. Since incubation of the product in Primatone RL‐supplemented acellular medium did not result in decreased sialylation, the negative effect of Primatone RL could not be attributed to extracellular desialylation of IFN‐γ by components of the peptone. In the fed‐batch mode, a culture utilizing a serum‐free feeding medium supplemented with Primatone RL demonstrated poorer sialylation than a similar culture not fed the peptone. The results of both the batch and fed‐batch experiments indicate that the adverse effect of the peptone was not due solely to ammonia accumulation. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 353–360, 1997.