Discovery of human monoclonal antibodies neutralizing serineproteases from Phage Display for antibotropic serum production

• Grant number: 24/01801-1
• Support Opportunities: Scholarships in Brazil - Master
• Start date: October 01, 2024
• End date: February 28, 2026
• Field of knowledge: Biological Sciences - Immunology
• Principal Investigator: Manuela Berto Pucca
• Grantee: Marcela Romanazzi Fernandes
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Associated scholarship(s): 25/11949-9 - Investigating the cross-recognition and clotting inhibition potencial of a new phage antibody against Moojase: a serine protease derived from Bothrops moojeni venom, BE.EP.MS

Abstract

The condition of snakebite is recognized as a neglected disease, exerting a significant impact on public health. Despite the existence of a specific treatment, antivenom, it presents several disadvantages, including adverse effects and high costs. In this context, this study proposes the discovery of monoclonal antibodies (scFv) with cross-reactivity, targeting serine proteases from Bothropic snakes.The development process involves initially conducting assays to evaluate the fibrinolytic and coagulant activity in solutions of fibrinogen and human plasma containing serine proteases. The selection of antibodies with high affinity and cross-reactivity will be carried out through the cross-panning method. Subsequently, the selected phage-antibodies will be amplified, and the phagemids will be recovered.The analysis of the chosen phage-antibodies will be performed through polyclonal ELISA, followed by the production of these antibodies using monoclonal ELISA. Positive clones will be sequenced, and fragments of human monoclonal antibodies will be produced and purified using the ELISA method.Additionally, in vitro neutralization assays will be conducted to evaluate the fibrinolytic and coagulant activity, while in vivo assays will address the hemorrhagic, nociceptive, and paw edema activities resulting from the presence of serine proteases. This study aims to significantly contribute to the development of effective therapies against snakebite, standing out for its comprehensive approach that spans from production to detailed functional characterization of the obtained monoclonal antibodies.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

NATTRODT, JANIO J. M.; BEZERRA-DE-FREITAS, VICTORIA A.; MERVAL, ANA PAULA S. S.; FILARDI, ELOISE T.; CERNI, FELIPE A.; DANTAS, DOMINGOS S. M.; LINS, ALYSSON B. M.; GALAN, LUIS E. B.; CARBONELL, ROBERTO C.; PUCCA, MANUELA B.. Case report: Clinical management of freshwater stingray wounds using negative pressure therapy. FRONTIERS IN MEDICINE, v. 12, p. 7-pg., 2025-03-10. (24/13258-0, 24/01801-1)

ROMANAZZI, MARCELA; FILARDI, ELOISE T. M.; PIRES, GEOVANNA M. M.; CERVEJA, MARCOS F.; MELO-DOS-SANTOS, GUILHERME; OLIVEIRA, ISADORA S.; FERREIRA, ISABELA G.; CERNI, FELIPE A.; SANTOS-FILHO, NORIVAL ALVES; MONTEIRO, WUELTON M.; et al. The Versatility of Serine Proteases from Brazilian Bothrops Venom: Their Roles in Snakebites and Drug Discovery. BIOMOLECULES, v. 15, n. 2, p. 25-pg., 2025-02-01. (23/15381-1, 23/11264-0, 24/08990-4, 24/01801-1, 24/13258-0, 20/13176-3)