Single Chain Variable Fragments Produced in Escherichia coli against Heat-Labile and Heat-Stable Toxins from Enterotoxigenic E. coli

PLoS One. 2015 Jul 8;10(7):e0131484. doi: 10.1371/journal.pone.0131484. eCollection 2015.

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.

Methods and findings: 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.

Conclusion: The developed recombinant scFvs against LT and ST constitute promising starting point for simple and cost-effective ETEC diagnosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Toxins / immunology*
  • Enterotoxigenic Escherichia coli / isolation & purification
  • Enterotoxigenic Escherichia coli / metabolism*
  • Enterotoxins / immunology*
  • Escherichia coli Proteins / immunology*
  • Molecular Sequence Data
  • Recombinant Proteins / isolation & purification
  • Single-Chain Antibodies / biosynthesis*
  • Single-Chain Antibodies / chemistry

Substances

  • Bacterial Toxins
  • Enterotoxins
  • Escherichia coli Proteins
  • Recombinant Proteins
  • Single-Chain Antibodies
  • heat stable toxin (E coli)
  • heat-labile enterotoxin, E coli

Grants and funding

This research was supported by grants 11/12928-2 from São Paulo Research Foundation (FAPESP) and from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq –301302/2013-8) to RMFP and by grants 04/08297-3 from São Paulo Research Foundation (FAPESP) to OHPR. CYO, CRFS, DDM and FBA were recipients of FAPESP fellowships (06/60258-8, 10/14659-6, 11/24111-0 and 11/22108-2, respectively). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.