Kishishita Shohei, Nishikawa Tomoko, Shinoda Yasuharu, Nagashima Hiroaki, Okamoto Hiroshi, Takuma Shinya, Aoyagi Hideki
Project Planning and Coordination Department, Project and Lifecycle Management Unit, Chugai Pharmaceutical Co., Ltd., 1-1 Nihonbashi-Muromachi 2-Chome, Chuo-ku, Tokyo 103-8324, Japan; Life Science and Bioengineering, Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1 Tennodai 1-Chome, Tsukuba, Ibaraki 305-8572, Japan.
API Process Development Department, Pharmaceutical Technology Division, Chugai Pharmaceutical Co., Ltd., 5-1 Ukima 5-Chome, Kita-ku, Tokyo 115-8543, Japan.
J Biosci Bioeng. 2015 Jun;119(6):700-5. doi: 10.1016/j.jbiosc.2014.10.028. Epub 2014 Nov 29.
During the production of therapeutic monoclonal antibodies (mAbs), not only enhancement of mAb productivity but also control of quality attributes is critical. Charge variants, which are among the most important quality attributes, can substantially affect the in vitro and in vivo properties of mAbs. During process development for the production of mAbs in a Chinese hamster ovary cell line, we have observed that an improvement in mAb titer is accompanied by an increase in the content of acidic charge variants. Here, to help maintain comparability among mAbs, we aimed to identify the process parameters that controlled the content of acidic charge variants. First, we used a Plackett-Burman design to identify the effect of selected process parameters on the acidic charge variant content. Eight process parameters were selected by using a failure modes and effects analysis. Among these, temperature shift was identified from the Plackett-Burman design as the factor most influencing the acidic charge variant content. We then investigated in more detail the effects of shift temperature and temperature shift timing on this content. The content decreased with a shift to a lower temperature and with earlier timing of this temperature shift. Our observations suggest that Plackett-Burman designs are advantageous for preliminary screening of bioprocess parameters. We report here for the first time that temperature downshift is beneficial for effective control of the acidic peak variant content.
在治疗性单克隆抗体（mAb）的生产过程中，提高mAb产量以及控制质量属性都至关重要。电荷变体是最重要的质量属性之一，会对mAb的体外和体内特性产生重大影响。在中国仓鼠卵巢细胞系生产mAb的工艺开发过程中，我们观察到mAb滴度的提高伴随着酸性电荷变体含量的增加。在此，为了帮助保持不同mAb之间的可比性，我们旨在确定控制酸性电荷变体含量的工艺参数。首先，我们使用Plackett-Burman设计来确定所选工艺参数对酸性电荷变体含量的影响。通过失效模式与效应分析选择了八个工艺参数。其中，温度变化从Plackett-Burman设计中被确定为对酸性电荷变体含量影响最大的因素。然后，我们更详细地研究了变化温度和温度变化时机对该含量的影响。随着温度降低以及温度变化时机提前，该含量会降低。我们的观察结果表明，Plackett-Burman设计有利于生物工艺参数的初步筛选。我们首次在此报告，降低温度有利于有效控制酸性峰变体含量。
