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

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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. © 2014 Chuang et al.

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Chuang, K. H., Hsieh, Y. C., Chiang, I. S., Chuang, C. H., Kao, C. H., Cheng, T. C., … Cheng, T. L. (2014). High-throughput sorting of the highest producing cell via a transiently protein-anchored system. PLoS ONE, 9(7). https://doi.org/10.1371/journal.pone.0102569

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