Journal
VACCINE
Volume 26, Issue 38, Pages 4969-4974Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.vaccine.2008.06.081
Keywords
ShRna; dhfr; IgG expression; CHO cells
Categories
Funding
- National Science Council
- VGHUST joint Research Program, Tsou's Foundation, Taiwan
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The dihydrofolate reductase (dhfr)/methotrexate (MTX) selection is a common method to conduct gene amplification in stable clones of Chinese hamster ovary (CHO) cells. We previously reported the use of a short hairpin RNA (shRNA) vector targeted to the dhfr gene resulted in improving the intracellular antigen expression in gene-amplified stable CHO cells [Hong, W.W., Wu, S.C., 2007. A novel RNA silencing vector to improve antigen expression and stability in Chinese hamster ovary cells. Vaccine 25(20), 4103-4111]. Here we investigated the use of the dhfr-targeted shRNA vector for immunoglobulin G(IgG) expression in gene-amplified stable CHO Cells. With the use of the dhfr-targeted shRNA vector, the gene-amplified CHO/dhFr(-) cells were found to increase IgG expression at 1.0 mu M MTX by more than 100% and to improve the genomic stability of IgG expression in MTX-free cultures by approximately 30%. The use of the dhfr-targeted shRNA vector can enhance the IgG expression in the gene-amplified stable CHO cells and uphold the IgG expression in MTX-free cultures. Utilizing the dhfr-targeted shRNA vector may provide an alternative way to maneuver CHO cell factories for IgG production in cultures. (C) 2008 Elsevier Ltd. All rights reserved.
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