Combined gene and environmental engineering offers a synergetic strategy to enhance r-protein production in Chinese hamster ovary cells.
Combined gene and environmental engineering offers a synergetic strategy to enhance r-protein production in Chinese hamster ovary cells.
复制标题
基因和环境工程相结合提供了一种增强中国仓鼠卵巢细胞中 r 蛋白产量的协同策略。
DOI：
10.1002/bit.28000
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发表时间：
2022
影响因子：
3.8
通讯作者：
Torres M
中科院分区：
文献类型：
--
作者：
Torres M
环境生长抑制条件（GIC）已广泛用于提高中国仓鼠卵巢（CHO）细胞的细胞比生产率（qP），最常见的是温度降档和丁酸钠（NaBu）处理。CHO细胞中的B淋巴细胞诱导成熟蛋白-1（BLIMP1）过表达也可抑制细胞生长并增加产物滴度和qP。鉴于相似的反应，本研究评价了BLIMP1表达、低温和NaBu处理对CHO细胞培养性能、细胞代谢和重组蛋白生产的单独和组合影响。正如预期的那样，所有三种干预措施都降低了细胞生长，将细胞阻滞在G1/G0细胞周期期，并增加了qP。然而，当考虑到细胞活力、重组基因表达和细胞代谢时，CHO细胞呈现出不同的反应，这表明BLIMP1和GIC产生更高生产率的分子位点存在差异。BLIMP1表达和GIC的组合协同作用以抑制细胞生长并最大化r蛋白产生，其中BLIMP1/NaBu条件导致产物滴度和qP的最显著改善。后一种条件也证明在高密度灌注培养中显著增加了产物产量（高达9.8 g免疫球蛋白G1 [IgG1]/L和2.2 g促红细胞生成素-Fc [EPO-Fc]/L）和qP（IgG1高达179 pg/细胞/天[pcd]，EPO-Fc高达30 pcd）。这些发现提供了关于BLIMP1和GIC的生产力提高效果的机制见解，以及它们在产生高生产力过程中的互补性。
Environmental growth‐inhibition conditions (GICs) have been used extensively for increasing cell‐specific productivity (qP) of Chinese hamster ovary (CHO) cells, with the most common being temperature downshift and sodium butyrate (NaBu) treatment. B lymphocyte‐induced maturation protein‐1 (BLIMP1) overexpression in CHO cells can also inhibit cell growth and increase product titers and qP. Given the similar responses, this study evaluated the individual and combined effects of BLIMP1 expression, low temperature, and NaBu treatment on culture performance, cell metabolism, and recombinant protein production of CHO cells. As expected, all three interventions decreased cell growth, arrested cells in G1/G0 cell cycle phase, and increased qP. However, CHO cells presented different responses when considering cell viability, recombinant gene expression, and cell metabolism that indicated differences in the molecular loci by which BLIMP1 and GICs generated higher productivities. Combinations of BLIMP1 expression and GICs acted synergistically to inhibit cell growth and maximize r‐protein production, with the BLIMP1/NaBu condition leading to the most significant improvements in product titers and qP. This latter condition also proved to substantially increase product yields (up to 9.8 g immunoglobulin G1 [IgG1]/L and 2.2 g erythropoietin‐Fc [EPO‐Fc]/L) and qP(up to 179 pg/cell/day [pcd] for IgG1 and 30 pcd for EPO‐Fc) in high‐density perfusion cultures. These findings offered mechanistic insights about the productivity‐enhancing effects of BLIMP1 and GICs, as well as their complementarity for generating highly productive processes.