High-throughput sorting of the highest producing cell via a transiently protein-anchored system

Kuo Hsiang Chuang, Yuan Chin Hsieh, I. Shiuan Chiang, Chih Hung Chuang, Chien Han Kao, Ta Chun Cheng, Yeng Tseng Wang, Wen Wei Lin, Bing Mae Chen, Steve R. Roffler, Ming Yii Huang, Tian Lu Cheng

研究成果: 雜誌貢獻文章同行評審

7 引文 斯高帕斯(Scopus)

摘要

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.

原文英語
文章編號e102569
期刊PLoS One
9
發行號7
DOIs
出版狀態已發佈 - 七月 18 2014

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

指紋 深入研究「High-throughput sorting of the highest producing cell via a transiently protein-anchored system」主題。共同形成了獨特的指紋。

引用此