| Autor: |
Chuang, Kuo-Hsiang, Hsieh, Yuan-Chin, Chiang, I-Shiuan, Chuang, Chih-Hung, Kao, Chien-Han, Cheng, Ta-Chun, Wang, Yeng-Tseng, Lin, Wen-Wei, Chen, Bing-Mae, Roffler, Steve R., Huang, Ming-Yii, Cheng, Tian-Lu |
| Předmět: |
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| Zdroj: |
PLoS ONE; Jul2014, Vol. 9 Issue 7, p1-7, 7p |
| Abstrakt: |
Developing a high-throughput method for the effecient selection of the highest producing cell is very important for the production of recombinant protein drugs. Here, we developed a novel transiently protein-anchored system coupled with fluorescence activated cell sorting (FACS) for the efficient selection of the highest producing cell. A furin cleavage peptide (RAKR) was used to join a human anti-epithelial growth factor antibody (αEGFR Ab) and the extracellular-transmembrane-cytosolic domains of the mouse B7-1 antigen (B7). The furin inhibitor can transiently switch secreted αEGFR Ab into a membrane-anchored form. After cell sorting, the level of membrane αEGFR Ab-RAKR-B7 is proportional to the amount of secreted αEGFR Ab in the medium. We further selected 23 αEGFR Ab expressing cells and demonstrated a high correlation (R2 = 0.9165) between the secretion level and surface expression levels of αEGFR Ab. These results suggested that the novel transiently protein-anchored system can easily and efficiently select the highest producing cells, reducing the cost for the production of biopharmaceuticals. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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