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Production of HlyA and CNF1 bacterial toxins and biopanning of recombinant antibodies

Grant number: 19/23686-1
Support Opportunities:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): January 30, 2020
Effective date (End): April 25, 2020
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal Investigator:Roxane Maria Fontes Piazza
Grantee:Bruna de Lucca Caetano
Supervisor: Sachdev Sidhu
Host 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:16/16188-7 - Recombinant antibodies against alpha-hemolysin and CNF1 toxins for the diagnosis and therapy of uropathogenic Escherichia coli, BP.DR


Uropathogenic Escherichia coli (UPEC) are the main cause of urinary tract infection (UTI). Among the virulence factors produced by UPEC, alpha-hemolysin (HlyA) and cytotoxic necrotizing factor 1 (CNF1) stand out. These toxins become good targets for diagnosis and therapeutic interventions, once there is a need for a rapid and sensitive diagnostic method, as well as new therapies, which allows greater responsiveness in the treatment of UTI, reducing the social and economic weight of this pathology. Thus, generation of recombinant toxin that enables the development of specific antibodies, used later on as biotechnological tools is the purpose of this work. The uses of antibodies against the HlyA and CNF1 toxins has been described in the literature, and have contributed to the development of new technologies for the production of human proteome-based antibody. They are the so-called synthetic or recombinant human antibodies (rAb) obtained from the construction of libraries using various technologies, among them the phage display technology. Since UPEC strains express both the HlyA and CNF1, obtaining rAbs capable of recognizing and neutralizing these toxins is a promising proposal for the diagnosis and treatment of UTI caused by UPEC. Bacterial toxins and recombinant antibodies will be obtained through molecular biology techniques in collaboration with Dr. Sachdev Sidhu, researcher at the The Donnelly Centre, University of Toronto, Canada, a laboratory integrated with a high throughput antibody research platform and applied to the generation of therapeutic grade antibodies against hundreds of antigens. (AU)

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