{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,25]],"date-time":"2026-07-25T17:35:58Z","timestamp":1785000958461,"version":"3.55.0"},"reference-count":42,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2008,12,2]],"date-time":"2008-12-02T00:00:00Z","timestamp":1228176000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2009,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Disulfide bonds play a critical role in the stabilization of the immunoglobulin \u03b2\u2010sandwich sandwich. Under reducing conditions, such as those that prevail in the cytoplasm, disulfide bonds do not normally form and as a result most antibodies expressed in that compartment (intrabodies) accumulate in a misfolded and inactive state. We have developed a simple method for the quantitative isolation of antibody fragments that retain full activity under reducing conditions from large mutant libraries. In\n                    <jats:italic>E. coli<\/jats:italic>\n                    , inactivation of the cysteine oxidoreductase DsbA abolishes protein oxidation in the periplasm, which leads to the accumulation of scFvs and other disulfide\u2010containing proteins in a reduced form. Libraries of mutant scFvs were tethered onto the inner membrane of\n                    <jats:italic>dsbA<\/jats:italic>\n                    cells and mutants that could bind fluorescently labeled antigen in the reducing periplasm were screened by\n                    <jats:italic>A<\/jats:italic>\n                    nchored\n                    <jats:italic>P<\/jats:italic>\n                    eriplasmic\n                    <jats:italic>Ex<\/jats:italic>\n                    pression (APEx; Harvey\n                    <jats:italic>et al<\/jats:italic>\n                    ., Proc Natl Acad Sci USA 2004;101:9193\u20139198.). Using this approach, we isolated scFv antibody variants that are fully active when expressed in the cytoplasm or when the four Cys residues that normally form disulfides are substituted by Ser residues.\n                  <\/jats:p>","DOI":"10.1002\/pro.31","type":"journal-article","created":{"date-parts":[[2009,2,10]],"date-time":"2009-02-10T12:54:48Z","timestamp":1234270488000},"page":"259-267","source":"Crossref","is-referenced-by-count":29,"title":["Engineering antibody fragments to fold in the absence of disulfide bonds"],"prefix":"10.1002","volume":"18","author":[{"given":"Min Jeong","family":"Seo","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ki Jun","family":"Jeong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clinton E.","family":"Leysath","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew D.","family":"Ellington","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brent L.","family":"Iverson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George","family":"Georgiou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2009,1,2]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1997.1457"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi980712q"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M707078200"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7799(98)01268-2"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-540-73259-4_15"},{"key":"e_1_2_6_7_2","first-page":"327","article-title":"The therapeutic potential of intrabodies in neurologic disorders: focus on Huntington and Parkinson diseases","volume":"20","author":"Messer A","year":"2006","journal-title":"Bio Drugs"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.2174\/092986706776872899"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.2174\/156652408783565595"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1996.0726"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1327.1999.00443.x"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.96.21.11723"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.98.1.75"},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M205264200"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2005.03.011"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2005.12.043"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2008.02.012"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2003.09.073"},{"key":"e_1_2_6_19_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.2001.4756"},{"key":"e_1_2_6_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(03)00530-8"},{"key":"e_1_2_6_21_2","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6750-7-81"},{"key":"e_1_2_6_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2005.05.004"},{"key":"e_1_2_6_23_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.micro.55.1.21"},{"key":"e_1_2_6_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2004.02.012"},{"key":"e_1_2_6_25_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0020333"},{"key":"e_1_2_6_26_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M705147200"},{"key":"e_1_2_6_27_2","unstructured":"HarveyBR(2003)Anchored Periplasmic Expression (APEx): a versatile technology for the flow cytometric selection of high affinity antibodies fromEshcerichia coliexpressed libraries. 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