Assessment of neutralizing capacity of antibodies raised in mice against a Crotalus durissus terrificus phosphodiesterase

Phosphodiesterases (PDE) belong to a super-family of enzymes that have multiple roles in the metabolism of nucleotides. These enzyme classes have played such a critical role as tools in the field of molecular biology, but their function is little known today. Here, we evaluated the humoral resp...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Alonso, María del Rosario, Leiva, Laura Cristina Ana, Gay, Claudia Carolina, Fusco, Luciano Sebastián
Formato: Reunión
Lenguaje:Inglés
Publicado: Sociedad Argentina de Investigación Clínica 2026
Materias:
Neutralizing capacity of antibodies
Crotalus durissus terrificus
Mice
Acceso en línea:http://repositorio.unne.edu.ar/handle/123456789/60052
Aporte de:
RIUNNE - Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE) de Universidad Nacional del Nordeste
id I48-R184-123456789-60052
record_format dspace
spelling I48-R184-123456789-600522026-02-13T11:52:38Z Assessment of neutralizing capacity of antibodies raised in mice against a Crotalus durissus terrificus phosphodiesterase Alonso, María del Rosario Leiva, Laura Cristina Ana Gay, Claudia Carolina Fusco, Luciano Sebastián Neutralizing capacity of antibodies Crotalus durissus terrificus Mice Phosphodiesterases (PDE) belong to a super-family of enzymes that have multiple roles in the metabolism of nucleotides. These enzyme classes have played such a critical role as tools in the field of molecular biology, but their function is little known today. Here, we evaluated the humoral response induced by a PDE from Crotalus durissus terrificus (CDT) venom used as antigen. The PDE of the Crotalus durissus terrificus venom (CDT-PDE) was purified by gel filtration chromatography (Sephadex G-75) and ion exchange chromatography (HiTrap Q-FF). The CDT-PDE activity was tested by chromogenic reaction with sodium salt of bis (p-nitrophenyl) phosphate and the purity was monitored by SDS-PAGE. BALB /c mice (n= 5) were immunized subcutaneously with 0.3, 0.6 and 1.2 μg of enzyme (CDT-PDE) on days 1, 15 and 30, respectively. Freund's complete adjuvant was used in the first immunization and incomplete adjuvant in the booster. On day 50, mice were sacrificed and plasma antibody titers were evaluated by ELISA and cross-reactions with Bothrops alternatus and B. diporus venoms were analyzed by dot blotting. Mice plasma showed a specific response to CDT-PDE and antibodies gave titers around 1/1600. About of dot blotting test, C.d.t venom and it’s PDE showed strong spots were noted in each case, but partial and negative reactions were noted on B. diporus and B. alternatus, respectively. The neutralization capacity of PDE- antibodies was 40 and 20 % when different amounts of serum were analyzed (36.66 μl; 18.33 μl). These results demonstrated that CDT-PDE produces a humoral response. PDE-CDT antibodies recognize the crotalic PDE and, to a lesser extent, the PDE present in the venom of B. diporus. These antibodies can be used as tools in the study of the effect of PDE on platelet aggregation, as well as in the purification of PDE by affinity chromatography to improve the performance in the isolation from CDT venom. 2026-02-13T10:47:51Z 2026-02-13T10:47:51Z 2019-11 Reunión Alonso, María del Rosario, et al., 2019. Assessment of neutralizing capacity of antibodies raised in mice against a Crotalus durissus terrificus phosphodiesteras. En: Reunión Anual de Sociedades de Biociencia 2019. LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica. LI Reunión Anual de la Asociación Argentina de Farmacología Experimental. XXI Reunión Anual de la Sociedad Argentina de Biología. XXXI Reunión Anual de la Sociedad Argentina de Protozoología. IX Reunión Anual de la Asociación Argentina de Nanomedicinas. VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio. Buenos Aires: Sociedad Argentina de Investigación Clínica; Sociedad de Farmacología Experimental; Sociedad Argentina de Biología; Sociedad Argentina de Protozoología; Asociación Argentina de Nanomedicinas; Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio; Histochemical Society, p. 284-284. 0025-7680 1669-9106 http://repositorio.unne.edu.ar/handle/123456789/60052 eng https://medicinabuenosaires.com/revistas/vol79-19/s4/vol79_s4.pdf openAccess http://creativecommons.org/licenses/by-nc-nd/2.5/ar/ application/pdf p. 284-284 application/pdf Sociedad Argentina de Investigación Clínica Sociedad de Farmacología Experimental Sociedad Argentina de Biología Sociedad Argentina de Protozoología Asociación Argentina de Nanomedicinas Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio Histochemical Society
institution Universidad Nacional del Nordeste
institution_str I-48
repository_str R-184
collection RIUNNE - Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE)
language Inglés
topic Neutralizing capacity of antibodies
Crotalus durissus terrificus
Mice
spellingShingle Neutralizing capacity of antibodies
Crotalus durissus terrificus
Mice
Alonso, María del Rosario
Leiva, Laura Cristina Ana
Gay, Claudia Carolina
Fusco, Luciano Sebastián
Assessment of neutralizing capacity of antibodies raised in mice against a Crotalus durissus terrificus phosphodiesterase
topic_facet Neutralizing capacity of antibodies
Crotalus durissus terrificus
Mice
description Phosphodiesterases (PDE) belong to a super-family of enzymes that have multiple roles in the metabolism of nucleotides. These enzyme classes have played such a critical role as tools in the field of molecular biology, but their function is little known today. Here, we evaluated the humoral response induced by a PDE from Crotalus durissus terrificus (CDT) venom used as antigen. The PDE of the Crotalus durissus terrificus venom (CDT-PDE) was purified by gel filtration chromatography (Sephadex G-75) and ion exchange chromatography (HiTrap Q-FF). The CDT-PDE activity was tested by chromogenic reaction with sodium salt of bis (p-nitrophenyl) phosphate and the purity was monitored by SDS-PAGE. BALB /c mice (n= 5) were immunized subcutaneously with 0.3, 0.6 and 1.2 μg of enzyme (CDT-PDE) on days 1, 15 and 30, respectively. Freund's complete adjuvant was used in the first immunization and incomplete adjuvant in the booster. On day 50, mice were sacrificed and plasma antibody titers were evaluated by ELISA and cross-reactions with Bothrops alternatus and B. diporus venoms were analyzed by dot blotting. Mice plasma showed a specific response to CDT-PDE and antibodies gave titers around 1/1600. About of dot blotting test, C.d.t venom and it’s PDE showed strong spots were noted in each case, but partial and negative reactions were noted on B. diporus and B. alternatus, respectively. The neutralization capacity of PDE- antibodies was 40 and 20 % when different amounts of serum were analyzed (36.66 μl; 18.33 μl). These results demonstrated that CDT-PDE produces a humoral response. PDE-CDT antibodies recognize the crotalic PDE and, to a lesser extent, the PDE present in the venom of B. diporus. These antibodies can be used as tools in the study of the effect of PDE on platelet aggregation, as well as in the purification of PDE by affinity chromatography to improve the performance in the isolation from CDT venom.
format Reunión
author Alonso, María del Rosario
Leiva, Laura Cristina Ana
Gay, Claudia Carolina
Fusco, Luciano Sebastián
author_facet Alonso, María del Rosario
Leiva, Laura Cristina Ana
Gay, Claudia Carolina
Fusco, Luciano Sebastián
author_sort Alonso, María del Rosario
title Assessment of neutralizing capacity of antibodies raised in mice against a Crotalus durissus terrificus phosphodiesterase
title_short Assessment of neutralizing capacity of antibodies raised in mice against a Crotalus durissus terrificus phosphodiesterase
title_full Assessment of neutralizing capacity of antibodies raised in mice against a Crotalus durissus terrificus phosphodiesterase
title_fullStr Assessment of neutralizing capacity of antibodies raised in mice against a Crotalus durissus terrificus phosphodiesterase
title_full_unstemmed Assessment of neutralizing capacity of antibodies raised in mice against a Crotalus durissus terrificus phosphodiesterase
title_sort assessment of neutralizing capacity of antibodies raised in mice against a crotalus durissus terrificus phosphodiesterase
publisher Sociedad Argentina de Investigación Clínica
publishDate 2026
url http://repositorio.unne.edu.ar/handle/123456789/60052
work_keys_str_mv AT alonsomariadelrosario assessmentofneutralizingcapacityofantibodiesraisedinmiceagainstacrotalusdurissusterrificusphosphodiesterase
AT leivalauracristinaana assessmentofneutralizingcapacityofantibodiesraisedinmiceagainstacrotalusdurissusterrificusphosphodiesterase
AT gayclaudiacarolina assessmentofneutralizingcapacityofantibodiesraisedinmiceagainstacrotalusdurissusterrificusphosphodiesterase
AT fuscolucianosebastian assessmentofneutralizingcapacityofantibodiesraisedinmiceagainstacrotalusdurissusterrificusphosphodiesterase
_version_ 1859884124279406592

Ejemplares similares