New strategies for recombinant antibodies generation against Stx1 and Stx2 toxins produced by Shiga toxin-producing E. coli

• Grant number: 13/03160-9
• Support type: Scholarships abroad - Research Internship - Doctorate
• Effective date (Start): August 01, 2013
• Effective date (End): January 31, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal researcher: Roxane Maria Fontes Piazza
• Grantee: Daniela Luz Hessel da Cunha
• Supervisor abroad: Sachdev S. Sidhu
• Home Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Research place: University of Toronto (U of T), Canada
• Associated to the scholarship: 10/20148-4 - Recombinant antibodies (scFv) against the EspB protein and Stx1, Stx2 toxins : preparation and characterization of these tools for the immunodiagnosis of enteropatogenic Escherichia coli (EPEC) and Shiga producing Escherichia coli (STEC), BP.DR

Abstract

Shiga toxin-producing Escherichia coli is responsible for hemorrhagic colitis and hemolytic-uremic syndrome, which aggravates the condition of the infected patients. The diagnosis of this pathogen is not included in routine laboratories in developing countries. The development of tools for detecting these toxins becomes very relevant to their diagnosis. Antibodies have proven to be an excellent paradigm for the design of high-affinity, protein-based binding reagents. In our laboratory, IgG antibodies can be obtained by two ways; polyclonal antibodies (pAbs), which have different affinities and specificities, being produced in, limited quantities. In contrast, monoclonal antibodies (mAbs) have homogeneity, high specificity, and are unlimitedly produced by specialty cell culture. In order to maintain homogeneity and specificity of the monoclonal antibodies, associated to a large-scale production with lower cost, the genetic engineering has been used to obtain recombinant antibodies such as scFv and Fab fragments which have the complementarity-determining regions (CDR). This project aimed apply different strategies for obtain the single-chain variable fragments against Stx1 and Stx2 toxins to be used as tools in the Shiga toxin-producing E. coli immunodiagnosis. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

SHIGA, E. A.; GUTH, B. E. C.; PIAZZA, R. M. F.; LUZ, D. Comparative analysis of rapid agglutination latex test using single-chain antibody fragments (scFv) versus the gold standard Vero cell assay for Shiga toxin (Stx) detection. Journal of Microbiological Methods, v. 175, AUG 2020. Web of Science Citations: 0.

LUZ, DANIELA; SHIGA, EMERSON A.; CHEN, GANG; QUINTILIO, WAGNER; ANDRADE, FERNANDA B.; MARANHAO, ANDREA Q.; CAETANO, BRUNA A.; MITSUNARI, THAIS; SILVA, MIRIAM A.; ROCHA, LETICIA B.; MORO, ANA M.; SIDHU, SACHDEV S.; PIAZZA, ROXANE M. F. Structural Changes in Stx1 Engineering Monoclonal Antibody Improves Its Functionality as Diagnostic Tool for a Rapid Latex Agglutination Test. ANTIBODIES, v. 7, n. 1 MAR 2018. Web of Science Citations: 0.

LUZ, DANIELA; CHEN, GANG; MARANHAO, ANDREA Q.; ROCHA, LETICIA B.; SIDHU, SACHDEV; PIAZZA, ROXANE M. F. Development and Characterization of Recombinant Antibody Fragments That Recognize and Neutralize In Vitro Stx2 Toxin from Shiga Toxin-Producing Escherichia coli. PLoS One, v. 10, n. 3 MAR 19 2015. Web of Science Citations: 11.