{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T22:49:37Z","timestamp":1759963777077,"version":"build-2065373602"},"reference-count":29,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2018,9,20]],"date-time":"2018-09-20T00:00:00Z","timestamp":1537401600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["17K19007","18H01792"],"award-info":[{"award-number":["17K19007","18H01792"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Journal"],"published-print":{"date-parts":[[2019,1]]},"abstract":"<jats:sec><jats:label\/><jats:p>As intracellular antibodies (intrabodies) are highly promising tools for drug discovery, an innovative antibody screening platform in mammalian cells was previously developed by a single\u2010chain Fv (scFv)\u2010c\u2010kit growth sensor, which successfully selected rabies nucleoprotein and phosphoprotein\u2010specific intrabodies from a synthetic scFv library. Since the scFv\u2010c\u2010kit growth sensor releases a growth signal after forming oligomers due to binding to an oligomeric antigen, it is critical to use a library which does not contain self\u2010oligomeric scFvs to avoid the off\u2010target signal of the growth sensor. Here, a novel method to eliminate self\u2010oligomeric scFvs directly in the cytoplasm of mammalian cells is presented. A suicide switch by fusing an scFv with a cell\u2010death signaling domain to eliminate scFv oligomers is developed. It is found that among four cell\u2010death signaling domains, a suicide switch by fusing scFv with Fas\u2010associated death domain (FADD) can selectively reduce oligomeric scFvs. Furthermore, the library after eliminating scFv oligomers results in higher efficiency in the intrabody selection platform with a growth sensor. Collectively, the scFv\u2010FADD suicide switch can be applied to eliminate oligomeric scFvs from a library, which can consequently improve the quality of intracellular scFv libraries and accelerate the discovery of intrabodies in the future.<\/jats:p><\/jats:sec>","DOI":"10.1002\/biot.201800350","type":"journal-article","created":{"date-parts":[[2018,9,1]],"date-time":"2018-09-01T06:24:34Z","timestamp":1535783074000},"source":"Crossref","is-referenced-by-count":5,"title":["A Suicide Switch Directly Eliminates Intracellular scFv Oligomers in the Cytoplasm of Mammalian Cells"],"prefix":"10.1002","volume":"14","author":[{"given":"Thuy Duong","family":"Nguyen","sequence":"first","affiliation":[{"name":"Department of Bioengineering, Graduate School of Engineering The University of Tokyo 7\u20103\u20101 Hongo, Bunkyo\u2010ku Tokyo 113\u20108656 Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Teruyuki","family":"Nagamune","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Graduate School of Engineering The University of Tokyo 7\u20103\u20101 Hongo, Bunkyo\u2010ku Tokyo 113\u20108656 Japan"},{"name":"Department of Chemistry and Biotechnology, Graduate School of Engineering The University of Tokyo 7\u20103\u20101 Hongo, Bunkyo\u2010ku Tokyo 113\u20108656 Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2301-6171","authenticated-orcid":false,"given":"Masahiro","family":"Kawahara","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Graduate School of Engineering The University of Tokyo 7\u20103\u20101 Hongo, Bunkyo\u2010ku Tokyo 113\u20108656 Japan"},{"name":"Department of Chemistry and Biotechnology, Graduate School of Engineering The University of Tokyo 7\u20103\u20101 Hongo, Bunkyo\u2010ku Tokyo 113\u20108656 Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2018,9,20]]},"reference":[{"key":"e_1_2_8_1_1","doi-asserted-by":"publisher","DOI":"10.4161\/19420862.2015.989042"},{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrc3930"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.nbt.2011.03.022"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1097\/ICU.0000000000000403"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1080\/17474086.2017.1283213"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.2471\/BLT.16.178061"},{"key":"e_1_2_8_7_1","first-page":"141","volume":"8","author":"Tanaka T.","year":"2014","journal-title":"Biologics"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1095-1110"},{"key":"e_1_2_8_9_1","first-page":"135","volume":"911","author":"Pellis M.","year":"2012","journal-title":"Methods Mol. 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