Production of a murine mAb against Bothrops alternatus and B. neuwiedi snake venoms and its use to isolate a thrombin-like serine protease fraction
Research article published in International journal of biological macromolecules (2022)
Abstract
Accidents with snakes from the genus Bothrops represent ~90 % of all snakebites in Brazil. Monoclonal antibodies (mAbs) targeting venom components can be important assets for treating envenoming syndromes, for developing diagnostic tests and for research purposes. Therefore, in this study, we aimed to generate murine mAbs against the antigenic mixture of Bothropic venoms traditionally used as immunogen to produce Bothropic antivenoms in Brazil. ELISA showed that one of the produced mAbs recognizes B. alternatus and B. neuwiedi venoms (mAb anti-Ba/Bn) specifically and Western Blot revealed that this mAb binds to a single protein band of molecular mass of ≈50 kDa. MAb anti-Ba/Bn inhibited the coagulant activity but was unable to neutralize hemorrhagic and phospholipase A2 activities caused by the B. neuwiedi venom. MAb anti-Ba/Bn was immobilized to Sepharose beads and used for immunoaffinity chromatography of B. neuwiedi venom. Proteolytic activity assays indicated that the immunoaffinity-purified fraction (BnF-Bothrops neuwiedi fraction) has a serine protease thrombin-like profile, which was supported by coagulability assays in mice. Bottom-up proteomic analysis confirmed the prevalence of serine proteases in BnF using label-free quantification. In conclusion, this work characterized a mAb with neutralizing properties against B. neuwiedi coagulant activity and demonstrates that immunoaffinity chromatography using mAbs can be a useful technique for purification of bioactive toxic proteins from Bothrops spp. snake venoms.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Resumen
Production of a murine mAb against Bothrops alternatus and B. neuwiedi snake venoms and its use to isolate a thrombin-like serine protease fraction.
Por qué esto importa para la hirudoterapia
Este estudio produjo un anticuerpo monoclonal murino (mAc anti-Ba/Bn) dirigido contra los venenos de las serpientes Bothrops alternatus y B. neuwiedi, y lo utilizó mediante cromatografía de inmunoafinidad para aislar una fracción de serina proteasa (BnF) con actividad coagulante tipo trombina, confirmada mediante ensayos de coagulabilidad en ratones y proteómica sin marcaje. El mAc neutralizó la actividad coagulante, pero no las actividades hemorrágica ni fosfolipasa A2 del veneno de B. neuwiedi. Para el ámbito de ASH, la conexión es indirecta: el estudio se refiere a enzimas de veneno de serpiente descritas como "tipo trombina", una clase de enzimas anticoagulantes/coagulantes conceptualmente paralela a las presentes en la saliva de la sanguijuela, pero no se involucran sanguijuelas, proteínas derivadas de sanguijuela ni hirudoterapia. La relevancia se limita a la enzimología comparativa de proteínas que interactúan con la trombina provenientes de una fuente biológica diferente, y el estudio es de naturaleza preclínica.
Citación
Production of a murine mAb against Bothrops alternatus and B. neuwiedi snake venoms and its use to isolate a thrombin-like serine protease fraction
Belo AA et al. · International journal of biological macromolecules, 2022
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Añadido a la biblioteca ASH: May 27, 2026 · Última actualización del sitio: 18 de junio de 2026