Voronina E V, Seregin Y A, Litvinova N A, Shvets V I, Shukurov R R
PHARMAPARK LLC, Bldg. 1, 8 Nauchny proezd, Moscow, Russian Federation.
M.V. Lomonosov Moscow State Academy of Fine Chemical Technology, Moscow, Russian Federation.
Springerplus. 2016 Sep 15;5(1):1584. doi: 10.1186/s40064-016-3213-2. eCollection 2016.
Recombinant monoclonal antibodies (mAbs) against tumor necrosis factor alpha are widely used in the biopharmaceutical therapy of autoimmune diseases. Currently, a large number of drugs based on these antibodies are available. Accordingly, the development of these products for the Russian market is an important goal. The aim of the current study is to describe the development of one such technology. CHO-DG44-derived cell lines producing mAb were developed using two strategies, one based on individual clones and the other based on cell pools. To obtain recombinant cell lines with highly amplified genes of interest, the clones underwent dihydrofolate reductase-mediated gene amplification. Using the best strategy for the selection and amplification of mAb-producing clones, we achieved the production of more than 1 g/L in small scale, non-optimized conditions.
抗肿瘤坏死因子α的重组单克隆抗体（mAb）广泛应用于自身免疫性疾病的生物制药治疗。目前，有大量基于这些抗体的药物可供使用。因此，将这些产品引入俄罗斯市场是一个重要目标。本研究的目的是描述一种此类技术的开发过程。利用两种策略开发了产生mAb的CHO-DG44衍生细胞系，一种基于单个克隆，另一种基于细胞池。为了获得具有高度扩增的目的基因的重组细胞系，对克隆进行了二氢叶酸还原酶介导的基因扩增。使用最佳策略选择和扩增产生mAb的克隆，我们在小规模、非优化条件下实现了超过1 g/L的产量。
