Meents Heiko, Enenkel Barbara, Werner Rolf G, Fussenegger Martin
Institute of Biotechnology, Swiss Federal Institute of Technology, ETH Zurich, CH-8093 Zurich.
Biotechnol Bioeng. 2002 Sep 20;79(6):619-27. doi: 10.1002/bit.10322.
We have engineered dihydrofolate reductase-deficient (dhfr(-)) Chinese hamster ovary (CHO)-DUKX B11 cells adapted for growth in serum-free suspension cultures for unlinked muristerone-inducible expression of the cyclin-dependent kinase inhibitor p27Kip1 and constitutive expression of the soluble intercellular adhesion molecule-1 (sICAM), a potent common cold therapeutic. Conditional overexpression of p27Kip1 resulted in a sustained G1-specific growth arrest of transgenic CHO-DUKX associated with up to fivefold-increased specific sICAM productivity. Herein we exemplify the implementation of controlled proliferation technology in a major biopharmaceutical production cell line that is compatible with key requirements for large-scale production procedures, including constitutive transgene expression and anchorage-independent growth in serum-free media.
我们构建了二氢叶酸还原酶缺陷型（dhfr(-)）中国仓鼠卵巢（CHO）-DUKX B11细胞，使其适应无血清悬浮培养生长，用于细胞周期蛋白依赖性激酶抑制剂p27Kip1的非连锁莫孕酮诱导表达以及可溶性细胞间粘附分子-1（sICAM，一种有效的普通感冒治疗药物）的组成型表达。p27Kip1的条件性过表达导致转基因CHO-DUKX细胞出现持续的G1期特异性生长停滞，同时特异性sICAM的生产率提高了五倍。在此，我们举例说明了在一种主要的生物制药生产细胞系中实施可控增殖技术，该技术符合大规模生产程序的关键要求，包括组成型转基因表达和在无血清培养基中不依赖贴壁生长。
