Y. Katakura et al., Productivity enhancement of recombinant protein in CHO cells via specific promoter activation by oncogenes, CYTOTECHNOL, 31(1), 1999, pp. 103-109
To construct a recombinant protein highly producing cell lines, we have pre
viously developed the Oncogene Activated Production (OAP) system by using B
HK-21 cells. Here we verified the availability of the OAP system in CHO cel
ls. We firstly generated `primed' ras amplified CHO cells, ras clone I, by
introducing human c-Ha-ras oncogene into CHO cells. This ras clone I enable
s quick and easy establishment of recombinant protein hyper producing cell
lines by introduction reporter gene of interest. Then we generated I13 by i
ntroducing human interleukin 6 (hIL-6) gene as a reporter gene, which showe
d enhanced productivity rate as compared to A7 established by conventional
method. Furthermore, we found that hIL-6 production level of I13 was slight
ly improved by raising the CO2 concentration from 5 to 8% possibly because
of the enhanced growth rate. We further introduced the E1A oncogene, which
has been shown to have a synergistic effect on the recombinant protein prod
uction of the ras-amplified BHK-21 cells, then evaluated the productivity.
When culture in 5% CO2 condition, only the slight effect can be seen. Howev
er when cultured in 8% CO2 condition, not only cell number, but also produc
tivity increased significantly, resulted in great augmentation of hIL-6 pro
duction, maximum production being 88.6 mu g/ml/3 days. This study demonstra
tes that recombinant protein production level reached commercially desirabl
e level by utilizing our OAP system in CHO cells and optimizing the culture
condition.