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

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 se...

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Autores principales:	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
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5dc196a6c5a943068d07557d4daf1b39
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spelling	oai:doaj.org-article:5dc196a6c5a943068d07557d4daf1b392021-11-25T06:08:00ZHigh-throughput sorting of the highest producing cell via a transiently protein-anchored system.1932-620310.1371/journal.pone.0102569https://doaj.org/article/5dc196a6c5a943068d07557d4daf1b392014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25036759/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Developing 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.Kuo-Hsiang ChuangYuan-Chin HsiehI-Shiuan ChiangChih-Hung ChuangChien-Han KaoTa-Chun ChengYeng-Tseng WangWen-Wei LinBing-Mae ChenSteve R RofflerMing-Yii HuangTian-Lu ChengPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 7, p e102569 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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 High-throughput sorting of the highest producing cell via a transiently protein-anchored system.
description	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.
format	article
author	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
author_facet	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
author_sort	Kuo-Hsiang Chuang
title	High-throughput sorting of the highest producing cell via a transiently protein-anchored system.
title_short	High-throughput sorting of the highest producing cell via a transiently protein-anchored system.
title_full	High-throughput sorting of the highest producing cell via a transiently protein-anchored system.
title_fullStr	High-throughput sorting of the highest producing cell via a transiently protein-anchored system.
title_full_unstemmed	High-throughput sorting of the highest producing cell via a transiently protein-anchored system.
title_sort	high-throughput sorting of the highest producing cell via a transiently protein-anchored system.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/5dc196a6c5a943068d07557d4daf1b39
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High-throughput sorting of the highest producing cell via a transiently protein-anchored system.

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