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
This study produced a murine monoclonal antibody (mAb anti-Ba/Bn) targeting Bothrops alternatus and B. neuwiedi snake venoms and used it via immunoaffinity chromatography to isolate a serine protease fraction (BnF) with thrombin-like coagulant activity, confirmed by coagulability assays in mice and label-free proteomics. The mAb neutralized the coagulant but not hemorrhagic or phospholipase A2 activities of B. neuwiedi venom. For ASH's domain, the connection is indirect: the study concerns snake venom enzymes described as 'thrombin-like,' a conceptually parallel anticoagulant/coagulant enzyme class to those found in leech saliva, but no leeches, leech-derived proteins, or hirudotherapy are involved. The relevance is limited to comparative enzymology of thrombin-interacting proteins from a different biological source, and the study is preclinical in nature.
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: June 18, 2026