دورية أكاديمية
Production of a murine mAb against Bothrops alternatus and B. neuwiedi snake venoms and its use to isolate a thrombin-like serine protease fraction.
| العنوان: | Production of a murine mAb against Bothrops alternatus and B. neuwiedi snake venoms and its use to isolate a thrombin-like serine protease fraction. |
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| المؤلفون: | Belo AA; Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil; Labtest Diagnóstica SA, Minas, Gerais, Brazil., Naves de Souza DL; Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil., de Melo-Braga MN; Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil., Lopes de Souza L; Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil., Molina Molina DA; Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil., Vaz de Melo PD; Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil; Labtest Diagnóstica SA, Minas, Gerais, Brazil., Larsen MR; Department of Biochemistry and Molecular Biology, University of Southern Denmark, Denmark., Guerra-Duarte C; Diretoria de Pesquisa e Desenvolvimento, Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil., Chávez-Olórtegui C; Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil. Electronic address: olortegi@icb.ufmg.br. |
| المصدر: | International journal of biological macromolecules [Int J Biol Macromol] 2022 Aug 01; Vol. 214, pp. 530-541. Date of Electronic Publication: 2022 Jun 23. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 7909578 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-0003 (Electronic) Linking ISSN: 01418130 NLM ISO Abbreviation: Int J Biol Macromol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Amsterdam : Elsevier Original Publication: Guildford, Eng., IPC Science and Technology Press. |
| مواضيع طبية MeSH: | Bothrops* , Crotalid Venoms*/chemistry, Animals ; Antibodies, Monoclonal ; Mice ; Proteomics ; Serine Proteases ; Snake Venoms ; Thrombin |
| مستخلص: | 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. (Copyright © 2022 Elsevier B.V. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Anticoagulant; Bothrops; Monoclonal antibody; Serine protease |
| المشرفين على المادة: | 0 (Antibodies, Monoclonal) 0 (Crotalid Venoms) 0 (Snake Venoms) EC 3.4.- (Serine Proteases) EC 3.4.21.5 (Thrombin) |
| تواريخ الأحداث: | Date Created: 20220626 Date Completed: 20220719 Latest Revision: 20220719 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1016/j.ijbiomac.2022.06.127 |
| PMID: | 35753516 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1879-0003 |
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| DOI: | 10.1016/j.ijbiomac.2022.06.127 |