Reprogramming of Chinese hamster ovary cells towards enhanced protein secretion.
Reprogramming of Chinese hamster ovary cells towards enhanced protein secretion.
复制标题
中国仓鼠卵巢细胞重编程以增强蛋白质分泌。
DOI：
10.1016/j.ymben.2021.12.004
复制
发表时间：
2022
影响因子：
8.4
通讯作者：
Torres M
中科院分区：
文献类型：
--
作者：
Torres M
中国仓鼠卵巢（CHO）细胞的分泌缺陷表型是高水平生产生物药物的主要限制，特别是对于那些具有复杂分子结构和翻译后修饰的生物药物。为了提高CHO细胞的分泌能力，我们最近设计了CHO细胞宿主，使其过表达BLIMP 1（CHOB），这是一种细胞工程策略，改变了细胞机制，并导致不同rprotein的产物产量和细胞特异性生产率显着提高。在这里，作为我们先前研究的后续，我们开发了新的CHO细胞宿主，它们共同过表达BLIMP 1和XBP 1 s（CHOBX），这两种转录因子共同驱动抗体产生浆细胞的专业分泌功能。我们发现，CHOBX细胞表现出比CHOB细胞更好的性能，对于重组IgG 1和“难以表达”的EPO-Fc融合蛋白具有更好的产物产率和细胞特异性生产率。CHOBX衍生的细胞系中的这些改善是由于BLIMP 1和XBP 1的协同共表达引起的一系列生理和代谢变化。首先，细胞生长受到抑制，细胞周期停滞在G1/G 0期，这些特征与BLIMP 1表达水平直接相关。其次，细胞增加了蛋白质翻译（整体和重组蛋白），扩大了内质网，提高了它们更有效地分泌蛋白质的能力。最后，细胞显示出有利于能量产生的代谢特征，具有明显的乳酸转换和增加的氨基酸消耗。这项研究强调了转录因子对CHO细胞重编程至所需表型的价值，为工程细胞提供了增强重组治疗蛋白生产的新功能。
The deficient secretory phenotype of Chinese hamster ovary (CHO) cells is a major limitation for high-level production of biopharmaceuticals, particularly for those with complex molecular architectures and post-translational modifications. To improve CHO cell secretory capacity, we recently engineered CHO cell hosts to overexpress BLIMP1 (CHOB), in a cell engineering strategy that transformed the cellular machinery and led to significantly higher product yields and cell-specific productivities for different rproteins. Here, as a follow-up to our previous study, we developed new CHO cell hosts that co-overexpress BLIMP1 and XBP1s ( CHOBX ), two transcription factors that together drive the professional secretory function of antibody-producing plasma cells. We found that the CHOBX cells presented an improved performance over that of CHOB cells, with better product yields and cell-specific productivities for a recombinant IgG1 and a ‘difficult-to-express’ EPO-Fc fusion protein. These improvements in the CHOBX-derived cell lines resulted from a series of physiological and metabolic changes due to the synergetic co-expression of BLIMP1 and XBP1s. Firstly, cells presented an inhibited cell growth and arrested cell cycle in G1/G0 phase, features that were directly linked to BLIMP1 expression levels. Secondly, cells increased protein translation (both overall and recombinant protein), expanded the endoplasmic reticulum and improved their capacity to secrete protein more effectively. Lastly, cells showed a metabolic profile favouring energy production, with a pronounced lactate switch and increased consumption of amino acids. This study highlights the value of transcription factors for reprogramming CHO cells towards a desired phenotype, offering the potential to engineer cells with new functionalities for enhanced manufacturing of recombinant therapeutic proteins.
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影响因子：
32.4
作者：
Shaffer, AL;Lin, KI;Staudt, LM
通讯作者：
Staudt, LM
DOI：
10.1007/978-1-4939-6972-2_9
发表时间：
2017-01-01
期刊：
HETEROLOGOUS PROTEIN PRODUCTION IN CHO CELLS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Gallagher, Clair;Kelly, Paul S.
通讯作者：
Kelly, Paul S.
DOI：
10.1073/pnas.0605562104
发表时间：
2007-02-06
影响因子：
11.1
作者：
Yan, Junli;Jiang, Jianming;Chin, Keh-Chuang
通讯作者：
Chin, Keh-Chuang
影响因子：
32.4
作者：
Shaffer, AL;Shapiro-Shelef, M;Staudt, LM
通讯作者：
Staudt, LM
影响因子：
2.9
作者：
Rodriguez, J;Spearman, M;Butler, M
通讯作者：
Butler, M