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Expression of an scFv antibody fragment in Nicotiana benthamiana and in vitro assessment of its neutralizing potential against the snake venom metalloproteinase BaP1 from Bothrops asper.
Gomes, Marinna; Alvarez, Maria Alejandra; Quellis, Leonardo Ramos; Becher, Melina Laguia; Castro, Juciane Maria de Andrade; Gameiro, Jacy; Caporrino, Maria Cristina; Moura-da-Silva, Ana Maria; de Oliveira Santos, Marcelo.
Afiliação
  • Gomes M; Laboratorio de Genética, Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Rua José Lourenço Kelmer, S/n - Martelos, Juiz de Fora, MG, 36036-330, Brazil.
  • Alvarez MA; CONICET-Universidade Maimónides (CEBBAD), Hidalgo 775, Lab 603, Buenos Aires, Argentina.
  • Quellis LR; Laboratorio de Genética, Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Rua José Lourenço Kelmer, S/n - Martelos, Juiz de Fora, MG, 36036-330, Brazil.
  • Becher ML; CONICET-Universidade Maimónides (CEBBAD), Hidalgo 775, Lab 603, Buenos Aires, Argentina.
  • Castro JMA; Laboratorio de Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Rua José Lourenço Kelmer, S/n - Martelos, Juiz de Fora, MG, 36036-330, Brazil.
  • Gameiro J; Laboratorio de Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Rua José Lourenço Kelmer, S/n - Martelos, Juiz de Fora, MG, 36036-330, Brazil.
  • Caporrino MC; Laboratorio de Imunopatologia, Instituto Butantan, Av. Vital Brazil, 1500, Butantã, CEP 05503-900 São Paulo, SP, Brazil.
  • Moura-da-Silva AM; Laboratorio de Imunopatologia, Instituto Butantan, Av. Vital Brazil, 1500, Butantã, CEP 05503-900 São Paulo, SP, Brazil.
  • de Oliveira Santos M; Laboratorio de Genética, Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Rua José Lourenço Kelmer, S/n - Martelos, Juiz de Fora, MG, 36036-330, Brazil. Electronic address: marcelo.santos@ufjf.edu.br.
Toxicon ; 160: 38-46, 2019 Mar 15.
Article em En | MEDLINE | ID: mdl-30802471
Human accidents with venomous snakes represent an overwhelming public health problem, mainly in rural populations of underdeveloped countries. Their high incidence and the severity of the accidents result in 81,000 to 138,000 deaths per year. The treatment is based on the administration of purified antibodies, produced by hyper immunization of animals to generate immunoglobulins (Igs), and then obtained by fractionating hyper immune plasma. The use of recombinant antibodies is an alternative to conventional treatment of snakebite envenoming, particularly the Fv fragment, named the single-chain variable fragment (scFv). We have produced recombinant single chain variable fragment scFv against the venom of the pit viper Bothrops asper at high levels expressed transiently and stably in transgenic plants and in vitro cultures that is reactive to BaP1 (a metalloproteinase from B. asper venom). The yield from stably transformed plants was significantly (p > 0.05) higher than the results in from transient expression. In addition, scFvBaP1 yields from systems derived from stable transformation were: transgenic callus 62 µg/g (±2); biomass from cell suspension cultures 83 µg/g (±0.2); culture medium from suspensions 71.75 mg/L (±6.18). The activity of scFvBaP1 was confirmed by binding and neutralization of the fibrin degradation induced by BnP1 toxins from B. neuwiedi and by Atroxlysin Ia from B. atrox venoms. In the present work, we demonstrated the potential use of plant cells to produce scFvBaP1 to be used in the future as a biotechnological alternative to horse immunization protocols to produce anti-venoms to be used in human therapy against snakebites.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Metaloendopeptidases / Planticorpos / Anticorpos de Cadeia Única Tipo de estudo: Guideline Limite: Animals Idioma: En Revista: Toxicon Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Brasil País de publicação: Reino Unido
Texto completo
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Metaloendopeptidases / Planticorpos / Anticorpos de Cadeia Única Tipo de estudo: Guideline Limite: Animals Idioma: En Revista: Toxicon Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Brasil País de publicação: Reino Unido