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Glycobiology, 2000, Vol. 10, No. 4 421-430
© 2000 Oxford University Press

Remodeling of sugar chain structures of human interferon-{gamma}

Kazuhiro Fukuta, Reiko Abe, Tomoko Yokomatsu, Naoko Kono, Mineko Asanagi, Fumio Omae, Mari Toba Minowa2, Makoto Takeuchi2 and Tadashi Makino1

Life Science Laboratory, Mitsui Chemicals Inc., 1144 Togo, Mobara, Chiba 297–0017, Japan and 2Central Laboratories for Key Technology, KIRIN Brewery Co., Ltd., 1–13–5 Fuku-ura, Kanazawa-ku, Yokohama 236–0004, Japan

Natural human interferon (IFN)-{gamma} has mainly biantennary complex-type sugar chains and scarcely has multiantennary structures. We attempted to remodel the sugar chain structures using IFN-{gamma} as a model glycoprotein. To obtain the branching glycoforms of IFN-{gamma}, we introduced the genes for GnT-IV (UDP-N-acetylglucosamine:{alpha}-1,3-D-mannoside ß-1,4-N-acetylglucosaminyltransferase) and/or GnT-V (UDP-N-acetylglucosamine:{alpha}-1,6-D-mannoside ß-1,6-N-acetylglucosaminyltransferase) into Chinese hamster ovary (CHO) cells producing human IFN-{gamma}. The parental CHO cells produced IFN-{gamma} with biantennary sugar chains mainly. When the GnT-IV activity was increased, triantennary sugar chains with a branch produced by GnT-IV increased up to 66.9% of the total sugar chains. When the GnT-V activity was increased, triantennary sugar chains with a corresponding branch increased up to 55.7% of the total sugar chains. When the GnT-IV and -V activities were increased at a time, tetraantennary sugar chains increased up to 56.2% of the total sugar chains. The proportion of these multiantennary sugar chains corresponded to the intracellular activities of GnT-IV and -V. What is more, lectin blot and flow cytometric analysis indicated that the multi-branch structure of the sugar chains was increased not only on IFN-{gamma}, one of the secretory glycoproteins, but also on almost CHO cellular proteins by introducing either or both of the GnT genes. The results suggest that the branching structure of sugar chains of glycoproteins could be controlled by cellular GnT-IV and GnT-V activities. This technology can produce glycoforms out of natural occurrence, which should enlarge the potency of glycoprotein therapeutics.

1 To whom correspondence should be addressed


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