Development of technology for culturing a cell line producing a single-domain antibody fused with the Fc fragment of human IgG1

D. S. Polyansky, E. I. Ryabova, A. A. Derkaev, N. S. Starkov, I. S. Kashapova, D. Shcheblyakov, A. P. Karpov, I. Esmagambetov
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Abstract

Objectives. To develop an effective technology for the cultivation of Chinese hamster ovary (CHO) cells stably producing GamP2C5 antibody which is a component I of the GamCoviMab candidate drug for emergency prevention and therapy of  infection caused by SARS-CoV-2 virus; to select optimal cultivation parameters and to scale this technology in production.Methods. The study was performed on CHO GamP2C5 (clone 78) cell culture, producing a single-domain antibody fused to the Fc fragment of human IgG1 GamP2C5. Different culture media and supplements were used. Cells were cultured in Erlenmeyer flasks, Biostat® RM 20 wave-mixed bioreactor, Ambr® 250 mini bioreactors, STR 200 stirred-tank bioreactor.Results. Using molecular-genetic and biotechnological methods, a stable clone producer of CHO GamP2C5 antibody, clone 78, was obtained. Then a technique was worked out for the cultivation of the obtained clone producer on different culture media. The most suitable cultivation regimes, culture media, and optimal supplements were selected. This technology was tested in laboratory conditions in a 10-L reactor, and then successfully scaled up for production at the MedGamal Branch of the Gamaleya National Research Center for Epidemiology and Microbiology.Conclusions. This study demonstrates the fundamental feasibility of developing and scaling up a culture technology, in order to produce a drug based on a modified single-domain antibody with virus neutralizing activity against different strains of SARS-CoV-2 virus.
开发培养产生与人类 IgG1 的 Fc 片段融合的单域抗体的细胞系的技术
目的开发一种培养中国仓鼠卵巢(CHO)细胞稳定生产 GamP2C5 抗体的有效技术,该抗体是用于紧急预防和治疗 SARS-CoV-2 病毒感染的 GamCoviMab 候选药物的 I 组份;选择最佳培养参数并将该技术规模化生产。研究在 CHO GamP2C5(克隆 78)细胞培养物上进行,生产融合了人类 IgG1 GamP2C5 的 Fc 片段的单域抗体。使用了不同的培养基和补充剂。细胞在 Erlenmeyer 烧瓶、Biostat® RM 20 波混合生物反应器、Ambr® 250 迷你生物反应器和 STR 200 搅拌槽生物反应器中培养。利用分子遗传学和生物技术方法,获得了 CHO GamP2C5 抗体的稳定克隆生产者克隆 78。然后,研究了在不同培养基上培养克隆生产者的技术。选出了最合适的培养方案、培养基和最佳补充剂。这项技术在实验室条件下的 10 升反应器中进行了测试,然后在加马勒亚国家流行病学和微生物学研究中心的 MedGamal 分部成功地扩大了生产规模。这项研究证明了开发和扩大培养技术的基本可行性,以便生产出一种基于改良的单域抗体的药物,这种抗体具有中和病毒的活性,可抵抗不同的 SARS-CoV-2 病毒株。
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