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Appl. Environ. Microbiol. doi:10.1128/AEM.02106-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Efficient antibody production with the suppression of O-mannosylation in yeast

Kirin Pharma Co., Ltd., CMC R&D Laboratories, Gunma, Japan; Kirin Pharma Co., Ltd., Production Planning, Tokyo, Japan; Kirin Holdings Co., Ltd., Central Laboratories for Frontier Technology, Gunma, Japan; Kirin Pharma Co., Ltd., Discovery Research Laboratories, Gunma, Japan; Daiichi Sankyo Co., Ltd., Process Technology Research Laboratories, Fukushima, Japan; Daiichi Sankyo Co., Ltd., Advanced Technology Research Laboratories, Tokyo, Japan; National Institute of Advanced Industrial Science and Technology (AIST), Ibaraki, Japan; Graduate School of Life and Environmental Science, University of Tsukuba, Ibaraki, Japan

^* To whom correspondence should be addressed. Email: jigami.yoshi{at}aist.go.jp.

	Abstract

When antibodies were expressed in a methylotrophic yeast Ogataea minuta, we found that abnormal O-mannosylation occurred in the secreted antibody. Yeast-specific O-mannosylation is initiated by the addition of mannose at serine (Ser) or threonine (Thr) residues in the endoplasmic reticulum (ER) via protein O-mannosyltransferase (Pmt) activity. To suppress the addition of O-linked sugar chains on antibodies, we examined the possibility of inhibiting Pmt activity by the addition of a Pmt inhibitor during cultivation. The Pmt inhibitor was found to partially suppress the O-mannosylation on the antibodies. Surprisingly, the suppression of O-mannosylation was associated with an increased amount of assembled antibody (H2L2) and enhanced the antigen-binding activity of the secreted antibody. In this study, we demonstrated the expression of human antibody in O. minuta and elucidated the relationship between O-mannosylation and antibody production in yeast.

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