Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells

Table 2 Summary of protein characteristics and post-translational modifications of pdFVII and rFVII from three cell lines

	pdFVII	CHOrFVII	HEK293rFVII	BHKrFVII
rFVII specific activity calculated from PT clotting values per mg protein (antigen) [1000 IU/mg]	Not done, theoretical specific activity 2.0 kIU/mg	2.0	2.3	1.8
Peptide map	Similar
γ-carboxylation of E35; i.e. abundance of 10^th Gla [%]	82 %	53 %	56 %	53 %
Presence of propeptide	Not detected by peptide mapping
Hydroxylation of D68 [%]	<1 %	9 %	7 %	15 %
O-glycosylation on S52 / S60; only three major identified types shown [%]	31 %: Glc(Xyl)₂ / Fuc	56 %: Glc(Xyl)₂ / Fuc	62 %: Glc(Xyl)₂ / Fuc	21 %: Glc(Xyl)₂ / Fuc
	27 %: GlcXyl / Fuc	7 %: GlcXyl₂	7 %: GlcXyl / Fuc	50 %: Glc / Fuc
	15 %: Glc / Fuc	14 %: Glc / Fuc	10 %: Glc / Fuc	12 %: Glc / -

Peptide mapping and the PTMs were done by LC-MS after tryptic digest (see Methods). The raw data for γ-carboxylation and O-glycosylation of all FVII isolates can be found in the addional PDF file provided in Additional file 1: Tables S1 and S3, respectively. Legend: Fuc Fucose, Gla γ-carboxy glutamic acid, Glc Glucose, IU International units, Xyl Xylose

ISSN: 1472-6750