人IL—6在CHO细胞表达及其实验室生产工艺的建立

本文以PKCR6质粒为载体,构建了人IL-6的次级克隆PKCR6-IL-6,并成功地转染了CHO细胞,获得高表达、稳定分泌人IL-6的转基因细胞,由此建立了实验室生产IL-6的基本工艺,即从方瓶过渡到转瓶,经转瓶高密度培养,CHO上清中IL-6含量可以提高近6倍,这为扩大规模生产奠定了一定的基础。

Expression of Human Interleukin-6 (IL-6) in CHO Cell Line and Establishment of Technique for IL-6 Production in Laboratory Scale

The secondary cloning vector PKCR6 carrying the human IL-6 cDNA was constructed with PKCR6 plasmid. The CHO cell line that expressed high level of IL-6 gene was suceessfully obtained, and the specificity and stability were both identified. In addition, the basic technique for IL-6 production in laboratory scale was also established, this technique involved the exchange of normal CHO adhesion cell culture in flask into suspension culture in stirring bottle. The experimental results showed that the quantity of IL-6 in Lhe supernatant of stirring bottle culture increased up to about 6 times than that in normal culture. It suggests that these reults might be contributed as the fundamentals to the production of IL-6 in large scale for clinical uses.