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ANALYSIS OF GLYCOSYLATION HETEROGENEITY IN IFN-¥ PRODUCED BY CHO CELLS DURING BATCH AND CONTINUOUS CULTURE.
Curling, E. M. A. Hayter, P. Tong, J. Jenkins, N. Salmon, I. Bull, A. T.
Curling, E. M. A.
Hayter, P.
Tong, J.
Jenkins, N.
Salmon, I.
Bull, A. T.
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Issue Date
1991
Submitted date
2024-02-28
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Abstract
The production of recombinant human interferon-x (IFN- X¥ ) by Chinese hamster ovary (CHO) cells in serum-free medium was heterogeneous with up to twelve molecular weight variants secreted. Using the enzyme N-glycanase to remove all the (N-linked) oligosaccharides or tunicamycin to inhibit glycosylation, it was found that the variation was due to both variable asparagine residue occupation (Asnyg and/or Asn- 100) by complex oligosaccharide and to truncation of the IFN-$ polypeptide due to proteolytic cleavage at the carboxy-terminal end. Using neuraminidase treatment, terminal sialic acid expression was found to be invariant. When the CHD cells were grown in two litre batch suspension culture in serum-free medium further heterogeneity was discovered, seen as a reproducible shift towards the secretion of underglycosylated IFN-Y with time. This phenomenon was independent of glucose concentration in the medium. The IFN- ¥ was produced during the exponential growth phase and declined in line with the cell growth rate and glucose uptake rates. In contrast, during glucose-limited chemostat culture at a constant dilution rate of 0.015h”l, the relative levels of each major glycosylation variant did not change significantly, although the proportion of fully glycosylated IFN-¥ was still less than that seen at the start of batch culture while non-glycosylated IEN- ¥ represented 15% of the total IFN- ¥ secreted.
Citation
Protein glycosylation, 269 - 274
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Book chapter
conference paper
conference paper
Language
en
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Series/Report no.
GBF monographs ; Volume 15
ISSN
0930-4320
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ISBN
1560811846
3527283676
3527283676
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Attribution-NonCommercial-ShareAlike 4.0 International