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To optimize microbial production of single-chain variable fragments (scFvs), we have chosen five model targets, 1) a hapten, Zearalenone (ZEN) mycotoxin, along with infectious agents 2) rabies virus, 3)\n                      <jats:italic>Propionibacterium acnes<\/jats:italic>\n                      , 4)\n                      <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                      , and a cancer cell 5) acute myeloid leukemia cell line (HL-60). The scFv binders were affinity selected from a non-immunized human phage display scFv antibody library and genetically fused to the N-terminus of emerald green fluorescent protein (EmGFP). The scFv-EmGFP fusion constructs were subcloned into an expression vector, under the control of T7 promoter, C-terminally tagged with hexa-histidine and expressed in different\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                      (\n                      <jats:italic>E. coli<\/jats:italic>\n                      ) hosts. This enabled the detection of cells that expressed the correct scFv-EmGFP fusion, termed fluorobody, via bright fluorescent signal in the cytoplasm. Among the three\n                      <jats:italic>E. coli<\/jats:italic>\n                      hosts tested, an engineered\n                      <jats:italic>E. coli<\/jats:italic>\n                      B strain called SHuffle B that promotes disulfide bond formation in the cytoplasm appeared to be the most appropriate host. The recombinant fluorobodies were well expressed (2\u20138\u00a0mg\/L), possessed the fluorescence property of EmGFP, and retained the ability to bind to their cognate targets. Their specific bindings were demonstrated by ELISA, fluorescence-linked immunosorbent assay (FLISA), flow cytometry, and fluorescent microscope imaging. The fluorobody expression platform in this study could be further adopted as a one-step immunostaining technique based on scFv, isolated from phage display library to numerous desired targets.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Key points<\/jats:title>\n                    <jats:p>\n                      \u2022\n                      <jats:italic>E. coli SHuffle express T7 is a suitable expression host for scFv-EmGFP (fluorobody)<\/jats:italic>\n                    <\/jats:p>\n                    <jats:p>\n                      \u2022\n                      <jats:italic>Only the clones harboring scFv-EmGFP plasmid will show bright fluorescent signal<\/jats:italic>\n                    <\/jats:p>\n                    <jats:p>\n                      \u2022\n                      <jats:italic>This platform can be used to produce fluorobodies for numerous purposes<\/jats:italic>\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Graphical abstract<\/jats:title>\n                  <\/jats:sec>","DOI":"10.1007\/s00253-023-12395-6","type":"journal-article","created":{"date-parts":[[2023,2,2]],"date-time":"2023-02-02T04:10:06Z","timestamp":1675311006000},"page":"1959-1970","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Production and applications of fluorobody from redox-engineered Escherichia coli"],"prefix":"10.1007","volume":"107","author":[{"given":"Witsanu","family":"Srila","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thae Thae","family":"Min","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thitima","family":"Sumphanapai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kuntalee","family":"Rangnoi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mehmet","family":"Berkmen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2674-2419","authenticated-orcid":false,"given":"Montarop","family":"Yamabhai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2023,2,2]]},"reference":[{"issue":"3","key":"12395_CR1","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1016\/j.jfda.2013.07.009","volume":"21","author":"N Anukul","year":"2013","unstructured":"Anukul N, Vangnai K, Mahakarnchanakul W (2013) Significance of regulation limits in mycotoxin contamination in Asia and risk management programs at the national level. 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