Glycosylation pattern of humanized IgG-like bispecific antibody produced by recombinant CHO cells.
The glycosylation pattern of a humanized anti-EGFRxanti-CD3 bispecific single-chain diabody with an Fc portion (hEx3-scDb-Fc) produced by recombinant Chinese hamster ovary cells was evaluated and compared with those of a recombinant humanized anti-IL-8 antibody (IgG1) and human serum IgG. N-Linked oligosaccharide structures were estimated by two-dimensional high-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. No sialylation was observed with purified hEx3-scDb-Fc and the anti-IL-8 antibody. From the analysis of neutral oligosaccharides, approximately more than 90% of the N-linked oligosaccharides of hEx3-scDb-Fc and the anti-IL-8 antibody were alpha-1,6-fucosylated. The galactosylated biantennary oligosaccharides comprise over 40% of the total N-linked oligosaccharides in both hEx3-scDb-Fc and the anti-IL-8 antibody. The fully galactosylated biantennary oligosaccharides from hEx3-scDb-Fc and the anti-IL-8 antibody accounted for only 10% of the N-linked; however, more than 20% of the N-linked oligosaccharides were fully galactosylated biantennary oligosaccharides in human serum IgG. The glycosylation pattern of hEx3-scDb-Fc was quite similar to that of the anti-IL-8 antibody.
Kim
Wook-Dong
WD
Department of Biotechnology, Graduate School of Engineering, Osaka University, Yamadaoka 2-1, Suita, Osaka, Japan.
Tokunaga
Miwako
M
Ozaki
Hiroyuki
H
Ishibashi
Takuya
T
Honda
Kohsuke
K
Kajiura
Hiroyuki
H
Fujiyama
Kazuhito
K
Asano
Ryutaro
R
Kumagai
Izumi
I
Omasa
Takeshi
T
Ohtake
Hisao
H
D000818:Animals; D018033:Antibodies, Bispecific / Q000378:metabolism*; D016466:CHO Cells; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D003412:Cricetulus; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D018033:Antibodies, Bispecific; D007074:Immunoglobulin G; D011994:Recombinant Proteins