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dc.contributor.authorCheng, Chiu-Minen_US
dc.contributor.authorTzou, Shey-Cherngen_US
dc.contributor.authorZhuang, Ya-Hanen_US
dc.contributor.authorHuang, Chien-Chiaoen_US
dc.contributor.authorKao, Chien-Hanen_US
dc.contributor.authorLiao, Kuang-Wenen_US
dc.contributor.authorCheng, Ta-Chunen_US
dc.contributor.authorChuang, Chih-Hungen_US
dc.contributor.authorHsieh, Yuan-Chinen_US
dc.contributor.authorTai, Ming-Hongen_US
dc.contributor.authorCheng, Tian-Luen_US
dc.date.accessioned2014-12-08T15:36:05Z-
dc.date.available2014-12-08T15:36:05Z-
dc.date.issued2014-05-13en_US
dc.identifier.issn1932-6203en_US
dc.identifier.urihttp://dx.doi.org/10.1371/journal.pone.0097367en_US
dc.identifier.urihttp://hdl.handle.net/11536/24424-
dc.description.abstractSingle-chain variable fragments (scFvs) serve as an alternative to full-length monoclonal antibodies used in research and therapeutic and diagnostic applications. However, when recombinant scFvs are overexpressed in bacteria, they often form inclusion bodies and exhibit loss of function. To overcome this problem, we developed an scFv secretion system in which scFv was fused with osmotically inducible protein Y (osmY), a bacterial secretory carrier protein, for efficient protein secretion. Anti-EGFR scFv (alpha EGFR) was fused with osmY (N- and C-termini) and periplasmic leader sequence (pelB) to generate alpha EGFR-osmY, osmY-alpha EGFR, and pelB-alpha EGFR (control), respectively. In comparison with the control, both the osmY-fused alpha EGFR scFvs were soluble and secreted into the LB medium. Furthermore, the yield of soluble alpha EGFR-osmY was 20-fold higher, and the amount of secreted protein was 250-fold higher than that of osmY-alpha EGFR. In addition, the antigenbinding activity of both the osmY-fused alpha EGFRs was 2-fold higher than that of the refolded pelB-alpha EGFR from inclusion bodies. Similar results were observed with alpha TAG72-osmY and alpha Her2-osmY. These results suggest that the N-terminus of osmY fused with scFv produces a high yield of soluble, functional, and secreted scFv, and the osmY-based bacterial secretion system may be used for the large-scale industrial production of low-cost alpha EGFRprotein.en_US
dc.language.isoen_USen_US
dc.titleFunctional Production of a Soluble and Secreted Single-Chain Antibody by a Bacterial Secretion Systemen_US
dc.typeArticleen_US
dc.identifier.doi10.1371/journal.pone.0097367en_US
dc.identifier.journalPLOS ONEen_US
dc.citation.volume9en_US
dc.citation.issue5en_US
dc.citation.epageen_US
dc.contributor.department生物科技學系zh_TW
dc.contributor.departmentDepartment of Biological Science and Technologyen_US
dc.identifier.wosnumberWOS:000336369200104-
dc.citation.woscount0-
Appears in Collections:Articles


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