Production and characterization of scFv recombinant antibodies against toxins from Bothrops and Crotalus venoms

• Grant number: 12/01028-3
• Support Opportunities: Regular Research Grants
• Start date: June 01, 2012
• End date: May 31, 2014
• Field of knowledge: Biological Sciences - Immunology - Immunochemistry
• Principal Investigator: Irene Fernandes
• Grantee: Irene Fernandes
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated researchers: Eliana Faquim de Lima Mauro ; Geraldo Santana Magalhães

Abstract

Our goal in this project is to produce and characterize antibodies containing the single chain Fragment variable - scFv - anti-toxins from snake venoms, viewing to understand their mechanisms of action and a possible therapeutic alternative for snake bites of the genus Crotalus and Bothrops.Accidents caused by snake venoms were included in the list of Neglected Tropical Diseases of WHO. In Brazil, according to the Ministry of Health, occurred in 2009, more than 22,700 accidents involving snakes. Of these, the genus Bothrops is involved in 90% and genus Crotalus in 8%. However, accidents by Crotalus have the highest fatality rate (1.9%). ScFv human antibodies, obtained by phage display, against crotoxin, main toxin of Crotalus venom and three scFv mutants generated by in silico studies, will be constructed and expressed in E. coli. Then, these will be purified and tested for their affinity (for surface plasmon resonance - Biacore) and for their ability to neutralize the phospholipase activity, myotoxicity and degranulation of mast cells induced by crotoxin. In addition, scFv antibodies derived from murine monoclonal antibody against a metalloproteinase from Bothrops venom will be also constructed, expressed and tested for its neutralizing ability against fibrinolytic activity, and inflammation induced by BnP1 (Bothrops neuwiedi). Antibodies with neutralizing activity can be used as complementary therapy to poisoning, because the current therapy, although very efficient in the neutralization of systemic effects, is not very efficient for the local effects induced by snake venoms. (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)

CASTRO, J. M. A.; OLIVEIRA, T. S.; SILVEIRA, C. R. F.; CAPORRINO, M. C.; RODRIGUEZ, D.; MOURA-DA-SILVA, A. M.; RAMOS, O. H. P.; RUCAVADO, A.; GUTIERREZ, J. M.; MAGALHAES, G. S.; et al. A neutralizing recombinant single chain antibody, scFv, against BaP1, A P-I hemorrhagic metalloproteinase from Bothrops asper snake venom. Toxicon, v. 87, p. 81-91, SEP 2014. (12/01028-3, 04/08297-3)