Table 1.

A-fucosylation of antibody expression cell lines derived from stable transfection of glycoengineered host cell lines. The host cell lines were transfected with a standard plasmid used for development of expression cell lines for a human antibody (mAb2 or mAb3), or co-transfected with an antibody and a GnT-III/Man-II expression plasmid. The antibody plasmid also contained the DHFR gene, and methotrexate was added to the medium as selective agent. After about three weeks of clone selection, the plates were screened for antibody expression by ELISA. Antibody expressing clones were further expanded into shaken 24-well or 6-well plates to perform a batch culture with a defined cell count at start. The antibody was purified from the cell culture supernatant after 7 days by protein A HPLC. The proportion of a-fucosylated glycans was measured by RP-HPLC or MALDI-TOF MS.

Host Cell Line		Antibody expression cell lines			A-fucosylated glycans [%]
Clone	GnT-III/Man-II transfections	Number of cell lines	Transfection	Culture format	Detection method	Average	Minimum	Maximum	Related Figure
17	1	61	mAb2	6-well	RP-HPLC	21	14	29	Figure S2.B
51	1	65	mAb2	6-well	RP-HPLC	21	12	33
54	1	78	mAb2	24-well	MALDI	25	11	40
64	1	54	mAb2	24-well	MALDI	31	20	44
255	1	53	mAb2	24-well	MALDI	25	14	39
260	1	59	mAb2	6-well	RP-HPLC	21	11	31
17–483	2	69	mAb2	6-well	MALDI	39	27	52	Fig. 4A
17–494	2	73	mAb2	6-well	MALDI	44	29	63
51–201	2	80	mAb2	6-well	MALDI	43	29	61
64–30	2	40	mAb2	6-well	MALDI	40	30	52
64–6024	2	53	mAb2	6-well	MALDI	35	25	43
64–6123	2	24	mAb2	6-well	MALDI	41	18	53
260–501	2	44	mAb2	6-well	MALDI	26	21	38
260–1513	2	12	mAb2	6-well	MALDI	39	29	54
17–494	2	24	mAb3	6-well	MALDI	49	35	61	Fig. 4BFig. 7C
17–494	2	23	mAb3 + GnT-III/Man-II	6-well	MALDI	48	32	67
64–30	2	24	mAb3	6-well	MALDI	43	27	53
64–30	2	21	mAb3 + GnT-III/Man-II	6-well	MALDI	46	37	58