Monday, July 30, 2007
P60
A novel RNA silencing vector to improve antigen expression and stability in Chinese hamster ovary cells
Suh-Chin Wu and Willy Hong. Institute of Biotechnology and Department of Life Science, National Tsing Hua University, 101, Section 2, Guangfu Road, Hsinchu, 30013, Taiwan
Chinese hamster ovary (CHO) cells and dihydrofolate reductase (dhfr)/methotrexate (MTX) gene amplification system are routinely used to generate stable producer CHO cell clones in biopharmaceutical industries. The present study proposes a novel method by the co-amplification of the silencing vector targeted to dhfr gene for improvements of selecting high-producing clones in dhfr-deficient and wild-type CHO cells. Using the silencing vector also resulted in improving the stability of the recombinant protein expression in the absence of MTX in the CHO/dhFr- and wild-type CHO cells. This new method is proposed to generate highly expressed stable cell clones of both dhfr-deficient and wild-type CHO cells for recombinant antigen production. Utilization of the silencing vector designed in this study can improve antigen expression through dhfr-directed gene amplification in other dhfr-competent cell lines for vaccine development.
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