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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Ozaki, Christiane Y. [1] ; Silveira, Caio R. F. [1] ; Andrade, Fernanda B. [1] ; Nepomuceno, Roberto [1] ; Silva, Anderson [1] ; Munhoz, Danielle D. [1] ; Yamamoto, Bruno B. [1] ; Luz, Daniela [1] ; Abreu, Patricia A. E. [1] ; Horton, Denise S. P. Q. [1] ; Elias, Waldir P. [1] ; Ramos, Oscar H. P. [2] ; Piazza, Roxane M. F. [1]
Total Authors: 13
[1] Inst Butantan, Bacteriol Lab, Sao Paulo, SP - Brazil
[2] CEA, iBiTecs, SIMOPRO, Gif Sur Yvette - France
Total Affiliations: 2
Document type: Journal article
Source: PLoS One; v. 10, n. 7 JUL 8 2015.
Web of Science Citations: 2

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. (AU)

FAPESP's process: 11/12928-2 - New challenge for the diagnosis of diarrheagenic Escherichia coli: recombinant antibody obtention against different virulence factors
Grantee:Roxane Maria Fontes Piazza
Support Opportunities: Regular Research Grants
FAPESP's process: 10/14659-6 - Obtention and characterization of recombinant antibodies (scFv) against heat-stable (st) toxin produced by Enterotoxigenic Escherichia coli (ETEC)
Grantee:Caio Raony Farina Silveira
Support Opportunities: Scholarships in Brazil - Master
FAPESP's process: 11/24111-0 - Expression and purification optimization of recombinant antibody (scFv) against heat-labile toxin (LT) of enterotoxigenic Escherichia coli (ETEC)
Grantee:Danielle Dias Munhoz
Support Opportunities: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 04/08297-3 - AbEvo - (Antibodies Evolution): intelligent system for the development of antibodies based on the structure of the antigen
Grantee:Oscar Henrique Pereira Ramos
Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE