Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA
- Autores
- López, Gisela Lumila; Leiva, Laura Cristina Ana; Fusco, Luciano Sebastian
- Año de publicación
- 2023
- Idioma
- inglés
- Tipo de recurso
- documento de conferencia
- Estado
- versión publicada
- Descripción
- Snake envenoming is a serious medical problem in tropical developing countries and antivenoms are the only effective therapy. Antisera are produced by immunizing horses with snake venom and adjuvant, usually Freund´s adjuvant. Botropic venoms contain metalloproteinases (SVMPs), responsible for the local effects of envenoming, such as hemorrhage, edema and myotoxicity as well as systemic bleeding. SVMPs represent around 34,2 % of the protein composition from B. diporus venom and causing lesions during the immunization of animals. In search of new processes, focused on animal welfare for the production immunobiologicals, and taking into account that the toxic action of SVMPs is inhibited by Na2-EDTA, in the present work, the immune response of animals inoculated with B.diporus venom blocked with Na2-EDTA was evaluated. For this proposal, the B. diporus venom (1,9 mg/mL) was blocked by Na2-EDTA (200 mM, B.dV/ Na2-EDTA) and used as antigen. Previously to the inoculation, the excess of chelate was removed by molecular exclusion chromatography (Sephadex G-25). Likewise, the venom without inhibitor (B.dV) received the same treatment and in both cases, the effective neutralization of SVMPs using azocasein as substrate was determined. Group of 5 CF-1 mice were immunized subcutaneously on days 0, 15 and 30 with B.dV or B.dV/ Na2-EDTA (7-30μg or 14-60 μg) emulsified with complete Freund?s Adjuvant and incomplete (booster). On days 14, 29, 37 blood samples were collected from the tip of the animals tail and day 45 the final bleeding and the separation of the different sera were performed. These were destined for immunoassays and neutralization assays for proteolytic, indirect hemolytic, and coagulant activity. The results of the enzyme-linked immunosorbent assay showed that both anti-B.dV and anti-B.dV/EDTA serum had high antibody titers (1/74.850 ? 1/186.150) at the end of the immunization protocol. Regarding the Western Blot, the anti-B.dV/EDTA serum recognized the main bands, corresponding to the venom proteins, in a similar way as the anti-B.dV. Additionally, the experimental sera produced showed neutralizing capacity of the main toxic activities tested in vitro. This result shows that Na2-EDTA does not affect the immunogenicity of proteins since animals immunized with B.dV/ Na2-EDTA respond to B.diporus venom in a similar way to animals immunized with venom without inhibitor.
Fil: López, Gisela Lumila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Leiva, Laura Cristina Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Fusco, Luciano Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
XXIII Annual Meeting of the Argentinean Biology Society
Buenos Aires
Argentina
Sociedad Argentina de Biología - Materia
-
Na2EDTA
ANTIVENOM
ANTIBODY - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/224955
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Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTALópez, Gisela LumilaLeiva, Laura Cristina AnaFusco, Luciano SebastianNa2EDTAANTIVENOMANTIBODYhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Snake envenoming is a serious medical problem in tropical developing countries and antivenoms are the only effective therapy. Antisera are produced by immunizing horses with snake venom and adjuvant, usually Freund´s adjuvant. Botropic venoms contain metalloproteinases (SVMPs), responsible for the local effects of envenoming, such as hemorrhage, edema and myotoxicity as well as systemic bleeding. SVMPs represent around 34,2 % of the protein composition from B. diporus venom and causing lesions during the immunization of animals. In search of new processes, focused on animal welfare for the production immunobiologicals, and taking into account that the toxic action of SVMPs is inhibited by Na2-EDTA, in the present work, the immune response of animals inoculated with B.diporus venom blocked with Na2-EDTA was evaluated. For this proposal, the B. diporus venom (1,9 mg/mL) was blocked by Na2-EDTA (200 mM, B.dV/ Na2-EDTA) and used as antigen. Previously to the inoculation, the excess of chelate was removed by molecular exclusion chromatography (Sephadex G-25). Likewise, the venom without inhibitor (B.dV) received the same treatment and in both cases, the effective neutralization of SVMPs using azocasein as substrate was determined. Group of 5 CF-1 mice were immunized subcutaneously on days 0, 15 and 30 with B.dV or B.dV/ Na2-EDTA (7-30μg or 14-60 μg) emulsified with complete Freund?s Adjuvant and incomplete (booster). On days 14, 29, 37 blood samples were collected from the tip of the animals tail and day 45 the final bleeding and the separation of the different sera were performed. These were destined for immunoassays and neutralization assays for proteolytic, indirect hemolytic, and coagulant activity. The results of the enzyme-linked immunosorbent assay showed that both anti-B.dV and anti-B.dV/EDTA serum had high antibody titers (1/74.850 ? 1/186.150) at the end of the immunization protocol. Regarding the Western Blot, the anti-B.dV/EDTA serum recognized the main bands, corresponding to the venom proteins, in a similar way as the anti-B.dV. Additionally, the experimental sera produced showed neutralizing capacity of the main toxic activities tested in vitro. This result shows that Na2-EDTA does not affect the immunogenicity of proteins since animals immunized with B.dV/ Na2-EDTA respond to B.diporus venom in a similar way to animals immunized with venom without inhibitor.Fil: López, Gisela Lumila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Leiva, Laura Cristina Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Fusco, Luciano Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaXXIII Annual Meeting of the Argentinean Biology SocietyBuenos AiresArgentinaSociedad Argentina de BiologíaTech Science Press2023info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/224955Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA; XXIII Annual Meeting of the Argentinean Biology Society; Buenos Aires; Argentina; 2022; 1-10327-95451667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.biologia.org.ar/wp-content/uploads/2023/09/Biocell-47-Supl-3-2023-47_Jornada-Anual-SAB-2022.pdfinfo:eu-repo/semantics/altIdentifier/url/https://www.biologia.org.ar/jornadas/jornadas-anteriores/Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:29:22Zoai:ri.conicet.gov.ar:11336/224955instacron: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:34982025-09-29 10:29:22.94CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA |
title |
Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA |
spellingShingle |
Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA López, Gisela Lumila Na2EDTA ANTIVENOM ANTIBODY |
title_short |
Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA |
title_full |
Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA |
title_fullStr |
Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA |
title_full_unstemmed |
Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA |
title_sort |
Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA |
dc.creator.none.fl_str_mv |
López, Gisela Lumila Leiva, Laura Cristina Ana Fusco, Luciano Sebastian |
author |
López, Gisela Lumila |
author_facet |
López, Gisela Lumila Leiva, Laura Cristina Ana Fusco, Luciano Sebastian |
author_role |
author |
author2 |
Leiva, Laura Cristina Ana Fusco, Luciano Sebastian |
author2_role |
author author |
dc.subject.none.fl_str_mv |
Na2EDTA ANTIVENOM ANTIBODY |
topic |
Na2EDTA ANTIVENOM ANTIBODY |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
Snake envenoming is a serious medical problem in tropical developing countries and antivenoms are the only effective therapy. Antisera are produced by immunizing horses with snake venom and adjuvant, usually Freund´s adjuvant. Botropic venoms contain metalloproteinases (SVMPs), responsible for the local effects of envenoming, such as hemorrhage, edema and myotoxicity as well as systemic bleeding. SVMPs represent around 34,2 % of the protein composition from B. diporus venom and causing lesions during the immunization of animals. In search of new processes, focused on animal welfare for the production immunobiologicals, and taking into account that the toxic action of SVMPs is inhibited by Na2-EDTA, in the present work, the immune response of animals inoculated with B.diporus venom blocked with Na2-EDTA was evaluated. For this proposal, the B. diporus venom (1,9 mg/mL) was blocked by Na2-EDTA (200 mM, B.dV/ Na2-EDTA) and used as antigen. Previously to the inoculation, the excess of chelate was removed by molecular exclusion chromatography (Sephadex G-25). Likewise, the venom without inhibitor (B.dV) received the same treatment and in both cases, the effective neutralization of SVMPs using azocasein as substrate was determined. Group of 5 CF-1 mice were immunized subcutaneously on days 0, 15 and 30 with B.dV or B.dV/ Na2-EDTA (7-30μg or 14-60 μg) emulsified with complete Freund?s Adjuvant and incomplete (booster). On days 14, 29, 37 blood samples were collected from the tip of the animals tail and day 45 the final bleeding and the separation of the different sera were performed. These were destined for immunoassays and neutralization assays for proteolytic, indirect hemolytic, and coagulant activity. The results of the enzyme-linked immunosorbent assay showed that both anti-B.dV and anti-B.dV/EDTA serum had high antibody titers (1/74.850 ? 1/186.150) at the end of the immunization protocol. Regarding the Western Blot, the anti-B.dV/EDTA serum recognized the main bands, corresponding to the venom proteins, in a similar way as the anti-B.dV. Additionally, the experimental sera produced showed neutralizing capacity of the main toxic activities tested in vitro. This result shows that Na2-EDTA does not affect the immunogenicity of proteins since animals immunized with B.dV/ Na2-EDTA respond to B.diporus venom in a similar way to animals immunized with venom without inhibitor. Fil: López, Gisela Lumila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina Fil: Leiva, Laura Cristina Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina Fil: Fusco, Luciano Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina XXIII Annual Meeting of the Argentinean Biology Society Buenos Aires Argentina Sociedad Argentina de Biología |
description |
Snake envenoming is a serious medical problem in tropical developing countries and antivenoms are the only effective therapy. Antisera are produced by immunizing horses with snake venom and adjuvant, usually Freund´s adjuvant. Botropic venoms contain metalloproteinases (SVMPs), responsible for the local effects of envenoming, such as hemorrhage, edema and myotoxicity as well as systemic bleeding. SVMPs represent around 34,2 % of the protein composition from B. diporus venom and causing lesions during the immunization of animals. In search of new processes, focused on animal welfare for the production immunobiologicals, and taking into account that the toxic action of SVMPs is inhibited by Na2-EDTA, in the present work, the immune response of animals inoculated with B.diporus venom blocked with Na2-EDTA was evaluated. For this proposal, the B. diporus venom (1,9 mg/mL) was blocked by Na2-EDTA (200 mM, B.dV/ Na2-EDTA) and used as antigen. Previously to the inoculation, the excess of chelate was removed by molecular exclusion chromatography (Sephadex G-25). Likewise, the venom without inhibitor (B.dV) received the same treatment and in both cases, the effective neutralization of SVMPs using azocasein as substrate was determined. Group of 5 CF-1 mice were immunized subcutaneously on days 0, 15 and 30 with B.dV or B.dV/ Na2-EDTA (7-30μg or 14-60 μg) emulsified with complete Freund?s Adjuvant and incomplete (booster). On days 14, 29, 37 blood samples were collected from the tip of the animals tail and day 45 the final bleeding and the separation of the different sera were performed. These were destined for immunoassays and neutralization assays for proteolytic, indirect hemolytic, and coagulant activity. The results of the enzyme-linked immunosorbent assay showed that both anti-B.dV and anti-B.dV/EDTA serum had high antibody titers (1/74.850 ? 1/186.150) at the end of the immunization protocol. Regarding the Western Blot, the anti-B.dV/EDTA serum recognized the main bands, corresponding to the venom proteins, in a similar way as the anti-B.dV. Additionally, the experimental sera produced showed neutralizing capacity of the main toxic activities tested in vitro. This result shows that Na2-EDTA does not affect the immunogenicity of proteins since animals immunized with B.dV/ Na2-EDTA respond to B.diporus venom in a similar way to animals immunized with venom without inhibitor. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023 |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/conferenceObject Reunión Journal http://purl.org/coar/resource_type/c_5794 info:ar-repo/semantics/documentoDeConferencia |
status_str |
publishedVersion |
format |
conferenceObject |
dc.identifier.none.fl_str_mv |
http://hdl.handle.net/11336/224955 Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA; XXIII Annual Meeting of the Argentinean Biology Society; Buenos Aires; Argentina; 2022; 1-1 0327-9545 1667-5746 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/224955 |
identifier_str_mv |
Evaluation of an experimental botropic antivenom produced with venom blocked with Na2-EDTA; XXIII Annual Meeting of the Argentinean Biology Society; Buenos Aires; Argentina; 2022; 1-1 0327-9545 1667-5746 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/url/https://www.biologia.org.ar/wp-content/uploads/2023/09/Biocell-47-Supl-3-2023-47_Jornada-Anual-SAB-2022.pdf info:eu-repo/semantics/altIdentifier/url/https://www.biologia.org.ar/jornadas/jornadas-anteriores/ |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.coverage.none.fl_str_mv |
Internacional |
dc.publisher.none.fl_str_mv |
Tech Science Press |
publisher.none.fl_str_mv |
Tech Science Press |
dc.source.none.fl_str_mv |
reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) |
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CONICET Digital (CONICET) |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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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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13.070432 |