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Recombinant antibodies (scFv-intimin) as a tool for early detection of diarrhea caused by enteropathogenic Escherichia coli (EPEC) Escherichia coli and Shiga toxin producing (STEC).

Grant number: 11/14361-0
Support type:Scholarships in Brazil - Master
Effective date (Start): May 01, 2012
Effective date (End): February 28, 2013
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal researcher:Roxane Maria Fontes Piazza
Grantee:Andressa Caravelli
Home Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

Diarrhea is a major cause of infant mortality, causing approximately 2 million deaths each year worldwide, mostly among children under 5 years old. Among these pathogens, Escherichia coli (E. coli) diarrheiogenic is considered an important etiologic agent of diarrheal diseases. One of the main virulence factors involved in the pathogenesis of enteropathogenic Escherichia coli (EPEC) and Shiga toxin producing Escherichia coli (STEC) is the intimin, an outer membrane protein that mediates the intimate adherence of bacteria to the epithelial cell. The detection of STEC and EPEC (typical and atypical) is the fundamental importance to defining the therapeutic in the E. coli infections, which is still the leading cause of acute diarrhea in children and adults in many developed and developing countries. Thus, antibodies are important tools in the detection of many pathogens. Recombinant DNA technology allows the development of recombinant antibody, maintaining or improve their functional properties. Currently it is possible to produce antibody fragments as recombinant proteins using different expression systems based on the structure of the single chain variable (single chain Fragment variable or scFv). The scFv is the smallest antibody fragment that still retains the ability to combine with the antigen, are composed of antibody molecule variable domains (V), without the constant domains (C). In this work we will build a synthetic gene, which will be cloned in expression vector PAE, transformed with in E. coli BL21 (DE3) plys strain and the protein will be induced, with aim to optimize the expression of anti-intimin recombinant antibodies that was able to recognize the conserved region of purified intimin and alfa intimin (Int388-667) in typical EPEC isolate. This important tool will be used to standardization of a diagnostic method simple and quick detection of EPEC and STEC.

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