Glycosylation pattern of humanized IgG-like bispecific antibody produced by recombinant CHO cells
Glycosylation pattern of humanized IgG-like bispecific antibody produced by recombinant CHO cells
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DOI：
10.1007/s00253-009-2152-z
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发表时间：
2010-01-01
影响因子：
5
通讯作者：
Ohtake, Hisao
中科院分区：
文献类型：
--
作者：
Kim, Wook-Dong;Tokunaga, Miwako;Ohtake, Hisao
评价了由重组中国仓鼠卵巢细胞产生的具有Fc部分的人源化抗EGFR x抗CD 3双特异性单链双抗体（hEx 3-scDb-Fc）的糖基化模式，并与重组人源化抗IL-8抗体（IgG 1）和人血清IgG的糖基化模式进行了比较。通过二维高效液相色谱法和基质辅助激光解吸/电离飞行时间质谱法估计N-连接寡糖结构。用纯化的hEx 3-scDb-Fc和抗IL-8抗体未观察到唾液酸化。根据中性寡糖的分析，hEx 3-scDb-Fc和抗IL-8抗体的约90%以上的N-连接寡糖是α-1，6-岩藻糖基化的。半乳糖基化双触角寡糖占hEx 3-scDb-Fc和抗IL-8抗体中总N-连接寡糖的40%以上。来自hEx 3-scDb-Fc和抗IL-8抗体的完全半乳糖基化的双触角寡糖仅占N-连接寡糖的10%;然而，在人血清IgG中，超过20%的N-连接寡糖是完全半乳糖基化的双触角寡糖。hEx 3-scDb-Fc的糖基化模式与抗IL-8抗体的糖基化模式非常相似。
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.