张存超,赵亮,陈飞,范里,王泽宋,谭文松

• 1:华东理工大学生物反应器工程国家重点实验室

摘要(Abstract)：

产物比生成速率是衡量细胞株表现的重要指标,研究产物比生成速率有差异的细胞株有助于深入理解此参数,进而为获得具有高产物比生成速率的细胞株提供借鉴。以表达单克隆抗体的中国仓鼠卵巢(CHO)细胞为研究对象,通过有限稀释法筛选获得了抗体比生产速率比对照提高57.8%的亚克隆。以此为基础,对其生理生化特征、葡萄糖代谢以及抗体轻重链基因表达量进行了详细的分析。此亚克隆在Hyclone SFM4CHO培养基批次培养中,最高活细胞密度比对照降低了68.6%,其在指数生长期细胞干重、细胞直径、胞内总蛋白含量及胞内总蛋白和细胞直径比与对照相比均有不同程度的增加,且其表达产物的轻重链基因和信使RNA相对含量均有所提高。代谢方面,此亚克隆的葡萄糖比消耗速率比对照增加了77.9%,指数生长期线粒体相对合成活性比对照增加了172.5%,有助于增加产物的翻译速率。通过研究对表达单克隆抗体的高产亚克隆生理生化特征、葡萄糖代谢以及抗体轻重链基因表达量的分析,可为深入理解细胞培养过程中产物比生成速率的提高提供借鉴,同时为高产细胞株的构建和筛选提供指导。

关键词(KeyWords)： 有限稀释法;中国仓鼠卵巢细胞;抗体比生产速率;葡萄糖代谢

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(21106045);; 上海高校青年教师培养资助计划(hlg11011)

作者(Author): 张存超,赵亮,陈飞,范里,王泽宋,谭文松

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