Weng Zhibing, Jin Jian, Shao ChunHua, Li Huazhong
School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
Process Science and Production Department, Sunshine GuoJian Pharmaceutical (Shanghai), Shanghai, China.
Cytotechnology. 2020 Apr;72(2):259-269. doi: 10.1007/s10616-020-00375-x. Epub 2020 Mar 31.
Over the past decade, global interest in the development of therapeutic monoclonal antibodies (mAbs) has risen rapidly. As therapeutic agents, antibodies have shown marked efficacy in combatting a range of cancers and immune diseases with high target specificity and low toxicity (Carla Lucia et al. in PLoS ONE 6:e24071, 2011; Donaghy in MAbs 8:659-671, 2016; Nasiri et al. in J Cell Physiol 9:6441-6457, 2018; Teo et al. in Cancer Immunol Immunother 61:2295-2309, 2012). Recent advances in cell culture technology, such as high-throughput clone screening, have facilitated antibody production at concentrations exceeding 10 g/L (Chen et al. in BMC Immunol 19:35, 2018; Huang et al. in Biotechnol Prog 26:1400-1410, 2010; Lu et al. in Biotechnol Bioeng 110:191-205, 2013; Singh et al. in Biotechnol Bioeng 113:698-716, 2016). As titers have improved, the industry has begun to focus on the adjustment of target antibody quality profiles to improve efficacy. Cell lines, culture media, and culture conditions impact protein quality (Van Beers and Bardor in Biotechnol J 7:1473-1484, 2012). Optimization of critical quality attributes (CQAs), such as charge variants, can be achieved through bioprocess development and is the preferred approach as changes to the cell line or growth media used is considered unfavorable by regulatory bodies (Gawlitzek et al. in Biotechnol Bioeng 103:1164-1175, 2009; Jordan et al. in Cytotechnology 65:31-40, 2013; Pan et al. in Cytotechnology 69:39-56, 2016). In this study, the effect of process control and ion supplementation on charge variants of mAbs produced by Chinese hamster ovary (CHO) cells was investigated. Results of this study demonstrated that the concentration of Zn, duration of culturing, and temperature affect charge variants of a given mAb. Under the optimum conditions of 3L bioreactors, the most significant was that Zn and temperature shift could further improve the quality of antibody. The main peak increased by 12%, and the acid peak decreased by 16%. At the same time, there was no significant loss of titer. This study provided supporting evidence for methods to improve charge variants arising during mAb production.
在过去十年中，全球对治疗性单克隆抗体（mAb）开发的兴趣迅速上升。作为治疗剂，抗体在对抗一系列癌症和免疫疾病方面显示出显著疗效，具有高靶点特异性和低毒性（卡拉·露西亚等人，《公共科学图书馆·综合》6：e24071，2011年；多纳吉，《单克隆抗体》8：659 - 671，2016年；纳西里等人，《细胞生理学杂志》9：6441 - 6457，2018年；张等人，《癌症免疫与免疫治疗》61：2295 - 2309，2012年）。细胞培养技术的最新进展，如高通量克隆筛选，已促进抗体产量超过10 g/L（陈等人，《BMC免疫学》19：35，2018年；黄等人，《生物技术进展》26：1400 - 1410，2010年；卢等人，《生物技术与生物工程》110：191 - 205，2013年；辛格等人，《生物技术与生物工程》113：698 - 716，2016年）。随着滴度的提高，该行业已开始专注于调整目标抗体质量概况以提高疗效。细胞系、培养基和培养条件会影响蛋白质质量（范·贝尔斯和巴多尔，《生物技术杂志》7：1473 - 1484，2012年）。关键质量属性（CQA）的优化，如电荷变体，可以通过生物工艺开发来实现，并且这是首选方法，因为监管机构认为改变所用的细胞系或生长培养基是不利的（高维茨克等人，《生物技术与生物工程》103：1164 - 1175，2009年；乔丹等人，《细胞技术》65：31 - 40，2013年；潘等人，《细胞技术》69：39 - 56，2016年）。在本研究中，研究了工艺控制和离子补充对中国仓鼠卵巢（CHO）细胞产生的mAb电荷变体的影响。本研究结果表明，锌浓度、培养持续时间和温度会影响给定mAb的电荷变体。在3L生物反应器的最佳条件下，最显著的是锌和温度变化可以进一步提高抗体质量。主峰增加了12%，酸性峰降低了16%。同时，滴度没有显著损失。本研究为改善mAb生产过程中出现的电荷变体的方法提供了支持证据。
