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...

Autores
Gomes, Marinna; Alvarez, Maria Alejandra; Quellis, Leonardo Ramos; Laguía Becher, Melina; Castro, Juciane Maria de Andrade; Gameiro, Jacy; Caporrino, Maria Cristina; Silva, Ana Maria Moura da; Santos, Marcelo de Oliveira
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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
Materia
BAP1
BOTHROPS
HETEROLOGOUS EXPRESSION
IN VITRO PLANT CULTURES
MOLECULAR FARMING
SCFV
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/118222
Acceder
id CONICETDig_c16f8bc831db8669f54d6bc7e4748960
oai_identifier_str oai:ri.conicet.gov.ar:11336/118222
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling 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 asperGomes, MarinnaAlvarez, Maria AlejandraQuellis, Leonardo RamosLaguía Becher, MelinaCastro, Juciane Maria de AndradeGameiro, JacyCaporrino, Maria CristinaSilva, Ana Maria Moura daSantos, Marcelo de OliveiraBAP1BOTHROPSHETEROLOGOUS EXPRESSIONIN VITRO PLANT CULTURESMOLECULAR FARMINGSCFVhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Human 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; BrasilFil: Alvarez, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides; ArgentinaFil: Quellis, Leonardo Ramos. Universidade Federal de Juiz de Fora; BrasilFil: Laguía Becher, Melina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Maimónides; ArgentinaFil: Castro, Juciane Maria de Andrade. Universidade Federal de Juiz de Fora; BrasilFil: Gameiro, Jacy. Universidade Federal de Juiz de Fora; BrasilFil: Caporrino, Maria Cristina. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Silva, Ana Maria Moura da. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Santos, Marcelo de Oliveira. Universidade Federal de Juiz de Fora; BrasilPergamon-Elsevier Science Ltd2019-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/118222Gomes, Marinna; Alvarez, Maria Alejandra; Quellis, Leonardo Ramos; Laguía Becher, Melina; Castro, Juciane Maria de Andrade; et al.; 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; Pergamon-Elsevier Science Ltd; Toxicon; 160; 3-2019; 38-460041-0101CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.toxicon.2019.02.011info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0041010119300522info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:19:55Zoai:ri.conicet.gov.ar:11336/118222instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 15:19:55.995CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv 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
title 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
spellingShingle 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
BAP1
BOTHROPS
HETEROLOGOUS EXPRESSION
IN VITRO PLANT CULTURES
MOLECULAR FARMING
SCFV
title_short 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
title_full 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
title_fullStr 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
title_full_unstemmed 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
title_sort 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
dc.creator.none.fl_str_mv Gomes, Marinna
Alvarez, Maria Alejandra
Quellis, Leonardo Ramos
Laguía Becher, Melina
Castro, Juciane Maria de Andrade
Gameiro, Jacy
Caporrino, Maria Cristina
Silva, Ana Maria Moura da
Santos, Marcelo de Oliveira
author Gomes, Marinna
author_facet Gomes, Marinna
Alvarez, Maria Alejandra
Quellis, Leonardo Ramos
Laguía Becher, Melina
Castro, Juciane Maria de Andrade
Gameiro, Jacy
Caporrino, Maria Cristina
Silva, Ana Maria Moura da
Santos, Marcelo de Oliveira
author_role author
author2 Alvarez, Maria Alejandra
Quellis, Leonardo Ramos
Laguía Becher, Melina
Castro, Juciane Maria de Andrade
Gameiro, Jacy
Caporrino, Maria Cristina
Silva, Ana Maria Moura da
Santos, Marcelo de Oliveira
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv BAP1
BOTHROPS
HETEROLOGOUS EXPRESSION
IN VITRO PLANT CULTURES
MOLECULAR FARMING
SCFV
topic BAP1
BOTHROPS
HETEROLOGOUS EXPRESSION
IN VITRO PLANT CULTURES
MOLECULAR FARMING
SCFV
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv 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
description 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.
publishDate 2019
dc.date.none.fl_str_mv 2019-03
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/118222
Gomes, Marinna; Alvarez, Maria Alejandra; Quellis, Leonardo Ramos; Laguía Becher, Melina; Castro, Juciane Maria de Andrade; et al.; 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; Pergamon-Elsevier Science Ltd; Toxicon; 160; 3-2019; 38-46
0041-0101
CONICET Digital
CONICET
url http://hdl.handle.net/11336/118222
identifier_str_mv Gomes, Marinna; Alvarez, Maria Alejandra; Quellis, Leonardo Ramos; Laguía Becher, Melina; Castro, Juciane Maria de Andrade; et al.; 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; Pergamon-Elsevier Science Ltd; Toxicon; 160; 3-2019; 38-46
0041-0101
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.toxicon.2019.02.011
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0041010119300522
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Pergamon-Elsevier Science Ltd
publisher.none.fl_str_mv Pergamon-Elsevier Science Ltd
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.22299

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