title: Single Chain Variable Fragments Produced in Escherichia coli against Heat-Labile and Heat-Stable Toxins from Enterotoxigenic E. coli.
abstract: BACKGROUND
  Diarrhea is a prevalent pathological condition frequently associated to the colonization of the small intestine by enterotoxigenic Escherichia coli (ETEC) strains, known to be endemic in developing countries. These strains can produce two enterotoxins associated with the manifestation of clinical symptoms that can be used to detect these pathogens. Although several detection tests have been developed, minimally equipped laboratories are still in need of simple and cost-effective methods. With the aim to contribute to the development of such diagnostic approaches, we describe here two mouse hybridoma-derived single chain fragment variable (scFv) that were produced in E. coli against enterotoxins of ETEC strains.
  
  
  RESULTS
  Recombinant scFv were developed against ETEC heat-labile toxin (LT) and heat-stable toxin (ST), from previously isolated hybridoma clones. This work reports their design, construction, molecular and functional characterization against LT and ST toxins. Both antibody fragments were able to recognize the cell-interacting toxins by immunofluorescence, the purified toxins by ELISA and also LT-, ST- and LT/ST-producing ETEC strains.
  
  
  CONCLUSIONS
  The developed recombinant scFvs against LT and ST constitute promising starting point for simple and cost-effective ETEC diagnosis.
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Christiane Y
authors.initials: CY
authors.lastname: Ozaki
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Caio R F
authors.initials: CR
authors.lastname: Silveira
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Fernanda B
authors.initials: FB
authors.lastname: Andrade
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Roberto
authors.initials: R
authors.lastname: Nepomuceno
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Anderson
authors.initials: A
authors.lastname: Silva
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Danielle D
authors.initials: DD
authors.lastname: Munhoz
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Bruno B
authors.initials: BB
authors.lastname: Yamamoto
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Daniela
authors.initials: D
authors.lastname: Luz
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Patrícia A E
authors.initials: PA
authors.lastname: Abreu
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Denise S P Q
authors.initials: DS
authors.lastname: Horton
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Waldir P
authors.initials: WP
authors.lastname: Elias
authors.affiliation: CEA, iBiTecs, SIMOPRO, Gif sur Yvette, France.
authors.forename: Oscar H P
authors.initials: OH
authors.lastname: Ramos
authors.affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, SP, Brasil.
authors.forename: Roxane M F
authors.initials: RM
authors.lastname: Piazza
mesh_terms: D000595:Amino Acid Sequence; D001427:Bacterial Toxins / Q000276:immunology*; D054307:Enterotoxigenic Escherichia coli / Q000302:isolation & purification / Q000378:metabolism*; D004768:Enterotoxins / Q000276:immunology*; D029968:Escherichia coli Proteins / Q000276:immunology*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000302:isolation & purification; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000737:chemistry
chemical_list: D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C034151:heat stable toxin (E coli); C027538:heat-labile enterotoxin, E coli