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Biotechnology and Applied Biochemistry (2002) 36, (133–140) (Printed in Great Britain)

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Changes in the overall extent of protein glycosylation by Chinese hamster ovary cells over the course of batch culture

Inn H. Y. Yuk and Daniel I. C. Wang¹

Key words: G₀/G₁ phase, lipid-linked oligosaccharide.

Abbreviations used: CHO, Chinese hamster ovary; CMW, chloroform/methanol/water (10:10:3, by vol.); IFN-g, interferon-g; LLO, lipid-linked oligosaccharide; TCA, trichloroacetic acid; tPA, tissue plasminogen activator.

¹To whom correspondence should be addressed(e-mail dicwang@mit.edu).

Glycosylation of recombinant proteins can change during the culture of animal cells. Since lipid-linked oligosaccharides (LLOs) are the carbohydrate donors in N-linked glycosylation, their availability is postulated to influence the extent of glycosylation. To test this hypothesis, relative LLO and glycosylation levels in Chinese hamster ovary (CHO) cells were monitored over the course of batch culture for corresponding changes. Radiolabelling studies reveal that throughout the length of culture, intracellular LLO levels remained within a 2-fold range and overall CHO protein glycosylation varied by less than 30%. The implications of these results and hypotheses to explain the findings are discussed.

Received 15 November 2001/1 May 2002; accepted 12 July 2001

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