Development of an Expression Vector Engineering Strategy Based on tDNA Insulator Element for the Stable Expression of Vascular Endothelial Growth Factor Receptor-Fc Fusion Protein.

Azam Rahimpour, Meysam Mosallaei, Farzad Pourghazi, Sayed Hassan Tabatabaee, Reyhaneh Hoseinpoor, Es'hagh Pourmaleki, Mohsen Soosanabadi
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Abstract

During the past decades, tremendous advances have occurred in manufacturing recombinant therapeutic proteins in Chinese hamster ovary (CHO) cells. Nevertheless, the production of stable high-producing cell lines has remained a major obstacle in the development process of the CHO cell line. It has been shown that genomic regulatory elements can promote cell line development efficiency by improving transgenes' productivity and stability. Such elements include insulators, ubiquitous chromatin opening elements, scaffold/matrix attachment regions, and antirepressors. In addition, tDNA elements are shown to act as insulators in mammalian cells. This study examines the effect of the tDNA insulator on stable expression of a vascular endothelial growth factor receptor-Fc fusion protein.

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基于tDNA绝缘子元件稳定表达血管内皮生长因子受体fc融合蛋白的表达载体工程策略的建立。
在过去的几十年里,在中国仓鼠卵巢(CHO)细胞中制造重组治疗蛋白取得了巨大的进展。然而,稳定高产细胞系的产生仍然是CHO细胞系发育过程中的主要障碍。研究表明,基因组调控元件可以通过提高转基因的产量和稳定性来促进细胞系的发育效率。这些元件包括绝缘体、无处不在的染色质开口元件、支架/基质附着区域和抗抑制因子。此外,tDNA元素在哺乳动物细胞中起绝缘体的作用。本研究探讨了tDNA绝缘子对血管内皮生长因子受体- fc融合蛋白稳定表达的影响。
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