Rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS).

Antibodies and their derivatives are the most important agents in therapeutics and diagnostics. Even after the significant progress in the technology for antibody screening from huge libraries, it takes a long time to isolate an antibody, which prevents a prompt action against the spread of a diseas...

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Autores principales:	Sung Sun Yim, Hyun Bae Bang, Young Hwan Kim, Yong Jae Lee, Gu Min Jeong, Ki Jun Jeong
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/32686144334b45a7bc883dbfe17b8458
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spelling	oai:doaj.org-article:32686144334b45a7bc883dbfe17b84582021-11-25T05:57:03ZRapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS).1932-620310.1371/journal.pone.0108225https://doaj.org/article/32686144334b45a7bc883dbfe17b84582014-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0108225https://doaj.org/toc/1932-6203Antibodies and their derivatives are the most important agents in therapeutics and diagnostics. Even after the significant progress in the technology for antibody screening from huge libraries, it takes a long time to isolate an antibody, which prevents a prompt action against the spread of a disease. Here, we report a new strategy for isolating desired antibodies from a combinatorial library in one day by repeated fluorescence-activated cell sorting (FACS). First, we constructed a library of synthetic human antibody in which single-chain variable fragment (scFv) was expressed in the periplasm of Escherichia coli. After labeling the cells with fluorescent antigen probes, the highly fluorescent cells were sorted by using a high-speed cell sorter, and these cells were reused without regeneration in the next round of sorting. After repeating this sorting, the positive clones were completely enriched in several hours. Thus, we screened the library against three viral antigens, including the H1N1 influenza virus, Hepatitis B virus, and Foot-and-mouth disease virus. Finally, the potential antibody candidates, which show K(D) values between 10 and 100 nM against the target antigens, could be successfully isolated even though the library was relatively small (∼ 10(6)). These results show that repeated FACS screening without regeneration of the sorted cells can be a powerful method when a rapid response to a spreading disease is required.Sung Sun YimHyun Bae BangYoung Hwan KimYong Jae LeeGu Min JeongKi Jun JeongPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 10, p e108225 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sung Sun Yim Hyun Bae Bang Young Hwan Kim Yong Jae Lee Gu Min Jeong Ki Jun Jeong Rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS).
description	Antibodies and their derivatives are the most important agents in therapeutics and diagnostics. Even after the significant progress in the technology for antibody screening from huge libraries, it takes a long time to isolate an antibody, which prevents a prompt action against the spread of a disease. Here, we report a new strategy for isolating desired antibodies from a combinatorial library in one day by repeated fluorescence-activated cell sorting (FACS). First, we constructed a library of synthetic human antibody in which single-chain variable fragment (scFv) was expressed in the periplasm of Escherichia coli. After labeling the cells with fluorescent antigen probes, the highly fluorescent cells were sorted by using a high-speed cell sorter, and these cells were reused without regeneration in the next round of sorting. After repeating this sorting, the positive clones were completely enriched in several hours. Thus, we screened the library against three viral antigens, including the H1N1 influenza virus, Hepatitis B virus, and Foot-and-mouth disease virus. Finally, the potential antibody candidates, which show K(D) values between 10 and 100 nM against the target antigens, could be successfully isolated even though the library was relatively small (∼ 10(6)). These results show that repeated FACS screening without regeneration of the sorted cells can be a powerful method when a rapid response to a spreading disease is required.
format	article
author	Sung Sun Yim Hyun Bae Bang Young Hwan Kim Yong Jae Lee Gu Min Jeong Ki Jun Jeong
author_facet	Sung Sun Yim Hyun Bae Bang Young Hwan Kim Yong Jae Lee Gu Min Jeong Ki Jun Jeong
author_sort	Sung Sun Yim
title	Rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS).
title_short	Rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS).
title_full	Rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS).
title_fullStr	Rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS).
title_full_unstemmed	Rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS).
title_sort	rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (facs).
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/32686144334b45a7bc883dbfe17b8458
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Rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS).

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