N-linked glycan structures of a mouse monoclonal antibody produced from tobacco BY2 suspension-cultured cells
N-linked glycan structures of a mouse monoclonal antibody produced from tobacco BY2 suspension-cultured cells
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DOI：
10.1263/jbb.101.212
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发表时间：
2006-03-01
影响因子：
2.8
通讯作者：
Seki, T
中科院分区：
文献类型：
--
作者：
Fujiyama, K;Misaki, R;Seki, T
将编码来自小鼠单克隆抗体的 H 链和 L 链的 cDNA 引入烟草 BY2 细胞中，并通过 HPLC、外切葡萄糖苷酶消化和基质辅助激光解吸/电离飞行时间 (MALDI-TOF) 质谱法的组合来分析植物产生的抗体的糖链结构。确定的聚糖结构为Man(5-6)GlcNAc(2) (22.3%)、GlcNAcMan(5)GlcNAc(2) (3.1%)、GlcNAcMan(3)FucXylGlcNAc(2) (24.4%)、GlcNAcMan(3)XylGlcNAc(2) (17.8%)、Man(3)FucXylGlcNAc(2) (24.3%) 和 Man(3)XylGlcNAc(2) (8.1%)。转基因悬浮培养细胞产生的抗体的主要聚糖结构含有典型的植物平分β(1,2)-木糖和α(1,3)-岩藻糖残基，表明重组抗体在晚期高尔基体中进行了翻译后修饰。
cDNA encoding H- and L-chains from a mouse monoclonal antibody was introduced into tobacco BY2 cells, and the resulting sugar chain structures of plant-produced antibodies were analyzed by a combination of HPLC, exoglucosidase digestion and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The glycan structures determined were Man(5-6)GlcNAc(2) (22.3%), GlcNAcMan(5)GlcNAc(2) (3.1%), GlcNAcMan(3)FucXylGlcNAc(2) (24.4%), GlcNAcMan(3)XylGlcNAc(2) (17.8%), Man(3)FucXylGlcNAc(2) (24.3%), and Man(3)XylGlcNAc(2) (8.1%). The major glycan structures of the antibodies produced by transgenic suspension-cultured cells contain typical plant bisecting beta(1,2)-xylose and alpha(1,3)-fucose residues, suggesting the posttranslational modification of a recombinant antibody in the late Golgi apparatus.