Asselbergs F A, Rahuel J, Cumin F, Leist C
Biotechnology Department, CIBA-GEIGY Ltd., Basle, Switzerland.
J Biotechnol. 1994 Feb 14;32(2):191-202. doi: 10.1016/0168-1656(94)90182-1.
A process was developed to produce recombinant human renin for X-ray analysis and enzyme inhibition studies. An expression vector containing a human prorenin cDNA and expressing a mouse dihydrofolate reductase selection marker was transfected into dhfr-minus Chinese hamster ovary cells. After selection of cell strains with an increased gene copy number with methotrexate, cultures of the recombinant cells were scaled-up in serum-free media. Major improvements in cellular productivity were achieved by using continuous suspension cultures with cell recycling instead of an adherent culture system or batch-mode suspension cultures. The recombinant zymogen prorenin was purified and preparatively activated with trypsin. Enzymatic properties of the recombinant active renin are described.
已开发出一种生产重组人肾素的方法，用于X射线分析和酶抑制研究。将含有人类肾素原cDNA并表达小鼠二氢叶酸还原酶选择标记的表达载体转染到二氢叶酸还原酶缺陷型中国仓鼠卵巢细胞中。在用甲氨蝶呤选择基因拷贝数增加的细胞株后，重组细胞培养物在无血清培养基中进行放大培养。通过使用带有细胞循环的连续悬浮培养，而非贴壁培养系统或分批式悬浮培养，细胞生产力得到了显著提高。重组酶原肾素原被纯化并用胰蛋白酶进行制备性激活。文中描述了重组活性肾素的酶学性质。
