Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/27522
Title: Inhibition of the Myotoxicity Induced by Bothrops jararacussu Venom and Isolated Phospholipases A2 by Specific Camelid Single- Domain Antibody Fragments.
Authors: Prado, Nidiane D. R.
Pereira, Soraya dos Santos.
Silva, Michele P. da
Morais, Michelle S. S.
Kayano, Anderson M.
Moreira-Dill, Leandro Soares.
Luiz, Marcos Barros.
Zanchi, Fernando Berton.
Fuly, André L.
Huacca, Maribel E. F.
Fernandes, Cleberson F.
Calderon, Leonardo de Azevedo.
Zuliani, Juliana Pavan.
Silva, Luiz H. Pereira da
Soares, Andreimar Martins.
Stabeli, Rodrigo Guerino.
Fernandes, Carla Freire Celedônio.
Affilliation: Fundação Oswaldo Cruz. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil.
Universidade Federal Fluminense. Rio de Janeiro, RJ, Brasil.
Universidade Federal de Rondônia. Porto Velho, RO, Brasil.
Empresa Brasileira de Pesquisa Agropecuária. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil. / Universidade Federal de Rondônia. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil. / Universidade Federal de Rondônia. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil. / Universidade Federal de Rondônia. Porto Velho, RO, Brasil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brasil. / Centro de Pesquisa em Medicina Tropical. Porto Velho, RO, Brasil.
Abstract: Antivenoms, produced using animal hyperimmune plasma, remains the standard therapy for snakebites. Although effective against systemic damages, conventional antivenoms have limited efficacy against local tissue damage. Additionally, the hypersensitivity reactions, often elicited by antivenoms, the high costs for animal maintenance, the difficulty of producing homogeneous lots, and the instability of biological products instigate the search for innovative products for antivenomtherapy. In this study, camelid antibody fragments (VHH) with specificity to Bothropstoxin I and II (BthTX-I and BthTX-II), two myotoxic phospholipases from Bothrops jararacussu venom, were selected from an immune VHH phage display library. After biopanning, 28 and 6 clones recognized BthTX-I and BthTX-II by ELISA, respectively. Complementarity determining regions (CDRs) and immunoglobulin frameworks (FRs) of 13 VHH-deduced amino acid sequences were identified, as well as the camelid hallmark amino acid substitutions in FR2. Three VHH clones (KF498607, KF498608, and KC329718) were capable of recognizing BthTX-I byWestern blot and showed affinity constants in the nanomolar range against both toxins. VHHs inhibited the BthTX-II phospholipase A2 activity, and when tested for cross-reactivity, presented specificity to the Bothrops genus in ELISA. Furthermore, two clones (KC329718 and KF498607) neutralized the myotoxic effects induced by B. jararacussu venom, BthTX-I, BthTX-II, and by a myotoxin from Bothrops brazili venom (MTX-I) inmice. Molecular docking revealed that VHH CDRs are expected to bind the C-terminal of both toxins, essential for myotoxic activity, and to epitopes in the BthTX-II enzymatic cleft. Identified VHHs could be a biotechnological tool to improve the treatment for snake envenomation, an important and neglected world public health problem.
Keywords: Myotoxicity
Bothrops jararacussu
Phospholipase A2
Antibody
Issue Date: 2016
Publisher: Plos One
Citation: SOARES, A. M. et al. Inhibition of the Myotoxicity Induced by Bothrops jararacussu Venom and Isolated Phospholipases A2 by Specific Camelid Single-Domain Antibody Fragments. PLoS One, v. 11, p.1-22 , 2016.
Copyright: open access
Appears in Collections:RO - Artigos de Periódicos



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