Effects of overexpression of X-box binding protein 1 on recombinant protein production in Chinese hamster ovary and NS0 myeloma cells
Effects of overexpression of X-box binding protein 1 on recombinant protein production in Chinese hamster ovary and NS0 myeloma cells
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DOI：
10.1002/bit.21562
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发表时间：
2008-01-01
影响因子：
3.8
通讯作者：
Chao, Sheng-Hao
中科院分区：
文献类型：
--
作者：
Ku, Sebastian C. Y.;Ng, Daphne T. W.;Chao, Sheng-Hao
X-box结合蛋白1（XBP-1）是细胞分泌途径和未折叠蛋白反应（UPR）的关键调节因子。研究表明，XBP-1的剪接形式XBP-1 S作为转录激活因子发挥作用，并上调与内质网蛋白分泌和生物合成相关的许多基因。由于普遍认为某些重组蛋白的生产受到宿主细胞分泌能力的限制，因此可以通过过表达XBP-1 S来实现蛋白生产的增加。在这项研究中，XBP-1 S对单克隆抗体（MAb），干扰素γ（IFN γ）和促红细胞生成素（EPO）的生产力的影响，在中国仓鼠卵巢（CHO）和NS 0细胞系。结果表明，XBP-1 S可能成为一个决定性因素，只有当重组蛋白的积累超过宿主细胞的分泌能力。在瞬时转染系统中，蛋白质分泌的瓶颈是实现，XBP-1 S的过表达提高蛋白质滴度高达2.5倍。相反，XBP-1 S的过表达对不表现出任何分泌瓶颈的稳定细胞系的蛋白质生产率没有可检测的影响。我们的结论是，XBP-1 S的过表达是一种有效的策略，在增强重组蛋白的生产时，宿主细胞的分泌途径是饱和的高水平的重组蛋白的合成。
X-box binding protein 1 (XBP-1) is a key regulator of the cellular secretory pathway and unfolded protein response (UPR). It has been shown that the spliced form of XBP-1, XBP-1S, functions as a transcription activator and up-regulates many genes associated with protein secretion and biosynthesis of endoplasmic reticula. Since the production of some recombinant proteins is widely believed to be limited by the secretory capacity of the host cell, an increase in protein production may be achieved by overexpressing XBP-1S. In this study, the effects of XBP-1S on the productivity of monoclonal antibody (MAb), interferon gamma (IFN gamma), and erythropoietin (EPO) are examined in Chinese hamster ovary (CHO) and NS0 cell lines. Results show that XBP-1S may become a determinative factor only when accumulation of recombinant proteins exceeds the secretory capacity of the host cell. In transient transfection systems where a bottleneck in protein secretion was achieved, overexpression of XBP-1S improved protein titers by up to 2.5-fold. In contrast, overexpression of XBP-1S had no detectable effects on protein productivity of stable cell lines that did not exhibit any secretory bottleneck. We conclude that overexpression of XBP-1S is an effective strategy in enhancing recombinant protein production when the secretory pathway of the host cell is saturated by high-level synthesis of recombinant proteins.