Fucose content of monoclonal antibodies can be controlled by culture medium osmolality for high antibody-dependent cellular cytotoxicity
Fucose content of monoclonal antibodies can be controlled by culture medium osmolality for high antibody-dependent cellular cytotoxicity
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DOI：
10.1007/s10616-011-9377-2
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发表时间：
2012-05-01
期刊：
影响因子：
2.2
通讯作者：
Hosoi, Shinji
中科院分区：
文献类型：
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作者：
Konno, Yoshinobu;Kobayashi, Yuki;Hosoi, Shinji
抗体依赖性细胞毒性（ADCC）取决于与单克隆抗体（MAb）结合的寡糖的岩藻糖含量。由于具有低岩藻糖含量的MAb表现出高ADCC活性，因此控制MAb的去岩藻糖基化水平（deFuc%）并分析影响deFuc%的因素非常重要。在本研究中，我们观察到大鼠杂交瘤细胞系YB 2/0中产生的单克隆抗体的deFuc%与培养基渗透压浓度呈负相关，r（2）值高达0.92。此外，无论用于调节渗透压摩尔浓度的化合物类型（NaCl、KCl、岩藻糖、果糖、肌酸或甘露醇）如何，在1至400 L范围内的培养规模下，deFuc%均表现出相同的相关性。我们通过在灌注和补料分批培养中保持恒定的培养基渗透压摩尔浓度成功地控制了MAb deFuc%。与这些观察结果一致，逆转录PCR分析显示，参与糖酵解，GDP-岩藻糖供应和岩藻糖转移在低渗条件下的基因转录减少。
Antibody-dependent cellular cytotoxicity (ADCC) is dependent on the fucose content of oligosaccharides bound to monoclonal antibodies (MAbs). As MAbs with a low fucose content exhibit high ADCC activity, it is important to control the defucosylation levels (deFuc%) of MAbs and to analyze the factors that affect deFuc%. In this study, we observed that the deFuc% was inversely related to culture medium osmolality for MAbs produced in the rat hybridoma cell line YB2/0, with r (2) values as high as 0.92. Moreover, deFuc% exhibited the same correlation irrespective of the type of compound used for regulating osmolality (NaCl, KCl, fucose, fructose, creatine, or mannitol) at a culture scale ranging from 1 to 400 L. We succeeded in controlling MAb deFuc% by maintaining a constant medium osmolality in both perfusion and fed-batch cultures. In agreement with these observations, reverse transcription PCR analyses revealed decreased transcription of genes involved in glycolysis, GDP-fucose supply, and fucose transfer under hypoosmotic conditions.