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

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Abstract

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.

Original languageEnglish
Article numbere102569
JournalPLoS One
Volume9
Issue number7
DOIs
Publication statusPublished - Jul 18 2014

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ASJC Scopus subject areas

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

Cite this

Chuang, K. H., Hsieh, Y. C., Chiang, I. S., Chuang, C. H., Kao, C. H., Cheng, T. C., Wang, Y. T., Lin, W. W., Chen, B. M., Roffler, S. R., Huang, M. Y., & Cheng, T. L. (2014). High-throughput sorting of the highest producing cell via a transiently protein-anchored system. PLoS One, 9(7), [e102569]. https://doi.org/10.1371/journal.pone.0102569