Decoupling Growth and Protein Production in CHO Cells: A Targeted Approach.
Decoupling Growth and Protein Production in CHO Cells: A Targeted Approach.
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DOI：
10.3389/fbioe.2021.658325
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发表时间：
2021
影响因子：
5.7
通讯作者：
Rosser SJ
中科院分区：
文献类型：
--
作者：
Donaldson JS;Dale MP;Rosser SJ
中国人卵巢细胞的分批补料培养已被用于生产大量的生物治疗剂，特别是单克隆抗体。然而，越来越多的下一代生物治疗剂，如双特异性抗体和融合蛋白，难以使用标准补料分批工艺表达。分离细胞生长和生物制品生产正在成为生物制造行业越来越期望的策略，特别是对于难以表达的产品。在不产生重组蛋白的情况下，使细胞生长至高细胞密度（生长期），然后诱导重组蛋白的表达并停止细胞增殖（生产期），通常通过将诱导型基因表达系统与增殖控制策略相结合。分离生长和生产阶段允许细胞资源更有效地用于生长或生产，改善生长特性并增强难以表达的蛋白质的生产。然而，目前的哺乳动物细胞增殖控制方法依赖于温度变化和化学试剂，这与许多非增殖途径相互作用，导致对产品质量和培养物活力的可变影响。合成生物学提供了一种替代方法，通过战略性地靶向增殖途径来阻止细胞生长，但在工业生物生产中基本上尚未使用。由于微生物解耦系统的最新发展和现有哺乳动物细胞工程工具的进步，我们建议重新审视解耦生长和生产的合成生物学方法。
Fed-batch cultures of Chinese Hamster Ovary cells have been used to produce high quantities of biotherapeutics, particularly monoclonal antibodies. However, a growing number of next-generation biotherapeutics, such as bi-specific antibodies and fusion proteins, are difficult to express using standard fed-batch processes. Decoupling cell growth and biotherapeutic production is becoming an increasingly desired strategy for the biomanufacturing industry, especially for difficult-to-express products. Cells are grown to a high cell density in the absence of recombinant protein production (the growth phase), then expression of the recombinant protein is induced and cell proliferation halted (the production phase), usually by combining an inducible gene expression system with a proliferation control strategy. Separating the growth and production phases allows cell resources to be more efficiently directed toward either growth or production, improving growth characteristics and enhancing the production of difficult to express proteins. However, current mammalian cell proliferation control methods rely on temperature shifts and chemical agents, which interact with many non-proliferation pathways, leading to variable impacts on product quality and culture viability. Synthetic biology offers an alternative approach by strategically targeting proliferation pathways to arrest cell growth but have largely remained unused in industrial bioproduction. Due to recent developments in microbial decoupling systems and advances in available mammalian cell engineering tools, we propose that the synthetic biology approach to decoupling growth and production needs revisiting.
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