Ishii Yoichi, Imamoto Yasufumi, Yamamoto Rie, Tsukahara Masayoshi, Wakamatsu Kaori
Bio-process Research and Development Laboratories, Kyowa Hakko Kirin Co., Ltd., 100-1 Hagiwara-machi, Takasaki-shi, Gunma 370-0013, Japan; Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu-shi, Gunma 376-8515, Japan.
Bio-process Research and Development Laboratories, Kyowa Hakko Kirin Co., Ltd., 100-1 Hagiwara-machi, Takasaki-shi, Gunma 370-0013, Japan.
J Biosci Bioeng. 2015 Apr;119(4):478-85. doi: 10.1016/j.jbiosc.2014.09.017. Epub 2014 Nov 7.
Achieving high productivity and quality is the final goal of therapeutic antibody development, but the productivity and quality of antibodies are known to be substantially dependent on the nature of the cell lines expressing the antibodies. We characterized two contrasting cell lines that produce trastuzumab, namely cell line A with a high titer and a low aggregate content and cell line B with a low titer and a high aggregate content to identify the causes of the differences. We observed the following differences: cell growth (A > B), proportion of defucosylated oligosaccharides on antibodies (A < B), and proportion of covalent antibody aggregates (A > B). Our results suggest that the high monoclonal antibody (mAb) titers in cell line A is associated with the high proliferation and is not caused by the lactate metabolism shift (switching from lactate production to net lactate consumption). Rather, these differences can be accounted for by the following: levels of tricarboxylic acid cycle intermediates (A > B), ammonium ion levels (A ≤ B), and oxidative stress (A > B).
实现高产量和高质量是治疗性抗体开发的最终目标，但众所周知，抗体的产量和质量在很大程度上取决于表达抗体的细胞系的性质。我们对两种产生曲妥珠单抗的对比鲜明的细胞系进行了表征，即高滴度和低聚集体含量的A细胞系以及低滴度和高聚集体含量的B细胞系，以确定差异的原因。我们观察到以下差异：细胞生长（A＞B）、抗体上去岩藻糖基化寡糖的比例（A＜B）以及共价抗体聚集体的比例（A＞B）。我们的结果表明，A细胞系中高单克隆抗体（mAb）滴度与高增殖相关，并非由乳酸代谢转变（从乳酸产生转变为净乳酸消耗）引起。相反，这些差异可由以下因素解释：三羧酸循环中间体水平（A＞B）、铵离子水平（A≤B）和氧化应激（A＞B）。
