Graduate School of Science and Technology, Gunma University, Gunma, Japan.
Bio Process Research and Development Laboratories, Production Division, Kyowa Kirin Co. Ltd., Takasaki, Gunma, Japan.
Biotechnol Prog. 2024 May-Jun;40(3):e3447. doi: 10.1002/btpr.3447. Epub 2024 Feb 28.
Chinese hamster ovary (CHO) cells are widely used for the industrial production of therapeutic monoclonal antibodies (mAbs). To meet the increasing market demands, high productivity, and quality are required in cell culture. One of the critical attributes of mAbs, from a safety perspective, is mAb fragmentation. However, methods for preventing mAbs fragmentation in CHO cell culture are limited. In this study, we observed that the antibody fragment content increased with increasing titers in fed-batch cultures for all three cell lines expressing recombinant antibodies. Adding copper sulfate to the culture medium further increased the fragment content, suggesting the involvement of reactive oxygen species (ROS) in the fragmentation process. Though antioxidants may be helpful to scavenge ROS, several antioxidants are reported to decrease the productivity of CHO cells. Among the antioxidants examined, we observed that the addition of catechin or (-)-epigallocatechin gallate to the culture medium prevented fragmentation content by about 20% and increased viable cell density and titer by 30% and 10%, respectively. Thus, the addition of catechins or compounds of equivalent function would be beneficial for manufacturing therapeutic mAbs with a balance between high titers and good quality.
中国仓鼠卵巢(CHO)细胞广泛用于治疗性单克隆抗体(mAb)的工业生产。为了满足不断增长的市场需求,细胞培养需要高生产力和高质量。从安全性角度来看,mAb 的一个关键属性是 mAb 片段化。然而,预防 CHO 细胞培养中 mAb 片段化的方法有限。在这项研究中,我们观察到在表达重组抗体的所有三种细胞系的分批补料培养中,抗体片段含量随着滴度的增加而增加。在培养基中添加硫酸铜进一步增加了片段含量,表明活性氧(ROS)参与了片段化过程。尽管抗氧化剂可能有助于清除 ROS,但据报道,几种抗氧化剂会降低 CHO 细胞的生产力。在所检查的抗氧化剂中,我们观察到向培养基中添加儿茶素或(-)-表没食子儿茶素没食子酸酯可分别将片段含量降低约 20%,并将活细胞密度和滴度分别提高 30%和 10%。因此,添加儿茶素或具有等效功能的化合物将有利于制造在高滴度和良好质量之间取得平衡的治疗性 mAb。