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
| Autor: | Jacy Gameiro, Juciane Maria de Andrade Castro, Ana Maria Moura-da-Silva, Marcelo de Oliveira Santos, María Alejandra Alvarez, Marinna Gomes, Maria C Caporrino, Melina Laguía Becher, Leonardo Ramos Quellis |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Otras Ciencias Biológicas
Bothrops asper HETEROLOGOUS EXPRESSION Nicotiana benthamiana Venom Toxicology MOLECULAR FARMING Plantibodies Microbiology law.invention purl.org/becyt/ford/1 [https] Ciencias Biológicas SCFV law Neutralization Tests Crotalid Venoms Tobacco Single-chain variable fragment Animals Bothrops BAP1 purl.org/becyt/ford/1.6 [https] IN VITRO PLANT CULTURES biology Antivenins Metalloendopeptidases biology.organism_classification Plants Genetically Modified Recombinant Proteins Snake venom BOTHROPS Recombinant DNA biology.protein Antibody CIENCIAS NATURALES Y EXACTAS Single-Chain Antibodies |
| Zdroj: | CONICET Digital (CONICET) Consejo Nacional de Investigaciones Científicas y Técnicas instacron:CONICET |
| Popis: | 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. Fil: Gomes, Marinna. Universidade Federal de Juiz de Fora; Brasil Fil: Alvarez, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides; Argentina Fil: Quellis, Leonardo Ramos. Universidade Federal de Juiz de Fora; Brasil Fil: Laguía Becher, Melina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides; Argentina Fil: Castro, Juciane Maria de Andrade. Universidade Federal de Juiz de Fora; Brasil Fil: Gameiro, Jacy. Universidade Federal de Juiz de Fora; Brasil Fil: Caporrino, Maria Cristina. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil Fil: Silva, Ana Maria Moura da. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil Fil: Santos, Marcelo de Oliveira. Universidade Federal de Juiz de Fora; Brasil |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|