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. 2018 Dec 10;11(2):350–372. doi: 10.1080/19420862.2018.1551044

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© 2018 The Author(s). Published with license by Taylor & Francis Group, LLC.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.

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Figure 1. — Generation of IgG1 glycoforms with homogeneous glycoforms. (a) IgG1 N-glycosylation schematics and abbreviations used in this study for elucidating structures of complex type biantennary N-glycan. Presence of a core fucose is indicated by F; Ax indicates a biantennary glycan structure with the number (x) of GlcNAc on the antenna; Gx indicates the number (x) of galactose on the antenna; Sx indicates the number (x) of sialic acid on the antenna. Fuc: Fucose, Man: Mannose, GlcNAc: N-Acetylglucosamine, Gal: Galactose, SA: Sialic Acid. (b) Each core fucosylated and non-fucosylated homogeneous glycoform was generated from the same CHO cell line with or without Kifunensine to produce starting mAbs, control (ctrl) and high-mannose (HM). Subsequent deglycosylation and transglycosylation performed to obtain resulting homogeneous glycoforms. Endoglycosidase EndoS2 from S. pyogenes and D184 mutant as glycosynthase recombinantly expressed from E. coli. Glycan oxazolines (oxa) were used as well-defined donor substrates (Figure S1).