Human monoclonal antibodies (scFv) discovery with cross-reactivity and pH-dependent to metalloproteases from Bothrops spp

• Grant number: 20/13176-3
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: October 01, 2021
• End date: January 01, 2026
• Field of knowledge: Biological Sciences - Pharmacology - Toxicology
• Principal Investigator: Eliane Candiani Arantes Braga
• Grantee: Isadora Sousa de Oliveira
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated scholarship(s): 22/08964-8 - Discovery of human monoclonal antibodies (scFv) with cross-reactivity and pH-dependent targeting metalloprotease from Bothrops spp., BE.EP.PD

Abstract

Snakebite accidents affect a large number of individuals worldwide, but are prominent in subtropical and tropical countries, causing a large number of deaths, sequelae and deformities. Thus, snakebite was again included in the Neglected Tropical Diseases (NTDs) list in 2017. In Brazil, these accidents are mostly caused by snakes from Bothrops genus (~68% in 2019), which is represented by 27 species distributed throughout the national territory. The venom of this genus is mainly composed of metalloproteases (~30-70%), phospholipases (PLA2, ~7-40%) and serine proteases (~2-24%), varying quantitatively between species. These components make the venoms hemorrhagic, proteolytic and coagulant, characteristics that are responsible for many of their local and systemic manifestations. The treatment for these envenomings is the administration of heterologous antivenom serum, in this case, antibotropic, or mixed sera. However, there are still several disadvantages regarding its use, since they can trigger hypersensitivity reactions, such as anaphylaxis and Serum Disease. Thus, new technologies for antivenom production have become essential for the antivenoms improvement, such as the phage display technique, which allows the selection of fully human antibodies against the most varied antigens. Indeed, this methodology has already been used to produce specific and neutralizing human monoclonal antibodies for different toxins from scorpions, bees and snakes. In the case of Bothrops snakes, only human monoclonal antibodies specific for PLA2 have so far been produced using this technique. However, several preclinical and clinical efforts are still needed so that these new monoclonal antibodies can be commercialized and used as therapy. This research project seeks the selection and production of human monoclonal antibodies of single-chain fragment variable (scFvs) type, through the phage display technique, capable of recognizing and neutralizing metalloproteases from botropic venoms. The project will also produce improved antibodies, that is, they should be pH-depending and cross-reactive for metalloproteases of different botropic species, constituting a new generation antibotropic serum. (AU)

Scientific publications

(26)

(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)

FERREIRA, ISABELA; OLIVEIRA, ISADORA; BORDON, KARLA; REIS, MOUZARLLEM; WIEZEL, GISELE; SANCHEZ, CAROLINE; SANTOS, LUISA; SANTOS-FILHO, NORIVAL; PUCCA, MANUELA; ANTUNES, LUSANIA; et al. Beyond Angiogenesis: The Multitasking Approach of the First PEGylated Vascular Endothelial Growth Factor (CdtVEGF) from Brazilian Rattlesnake Venom. TOXINS, v. 15, n. 8, p. 21-pg., 2023-08-01. (20/13176-3, 21/11936-3, 17/00586-6, 19/10173-6)

DE OLIVEIRA NEVES, VIVIANO GOMES; DE MELLO, MARCELA M. BLASCKE; RODRIGUES, DANIEL; PERNOMIAN, LAENA; DE OLIVEIRA, ISADORA SOUSA; PARENTE, JULIANA M.; ARANTES, ELIANE CANDIANI; TOSTES, RITA C.; CASTRO, MICHELE M.. Type I collagen proteolysis by matrix metalloproteinase-2 contributes to focal adhesion kinase activation and vascular smooth muscle cell proliferation in the aorta in early hypertension. VASCULAR PHARMACOLOGY, v. 152, p. 9-pg., 2023-10-01. (19/10173-6, 20/13176-3, 19/09174-8)

OLIVEIRA, ISADORA S.; PUCCA, MANUELA B.; CERNI, FELIPE A.; VIEIRA, SAMUEL; SACHETT, JACQUELINE; DE FARIAS, ALTAIR SEABRA; LACERDA, MARCUS; MURTA, FELIPE; BAIA-DA-SILVA, DJANE; ROCHA, THIAGO AUGUSTO HERNANDES; et al. Snakebite envenoming in Brazilian children: clinical aspects, management and outcomes. JOURNAL OF TROPICAL PEDIATRICS, v. 69, n. 2, p. 13-pg., 2023-02-06. (20/13176-3)

FRANCO, MICHELLE V. S.; ALEXANDRE-SILVA, GABRIEL M.; OLIVEIRA, ISADORA S.; SANTOS, POLIANA L.; SANDRI, ELISEU A.; CERNI, FELIPE A.; PUCCA, MANUELA B.. Physical and social consequences of snakebites in the Yanomami indigenous community, Brazil: Report of two cases. Toxicon, v. 214, p. 2-pg., 2022-05-26. (20/13176-3)

ISADORA SOUSA DE OLIVEIRA; LARISSA SOARES CARDOSO; ISABELA GOBBO FERREIRA; GABRIEL MELO ALEXANDRE-SILVA; BEATRIZ DE CÁSSIA DA SILVA JACOB; FELIPE AUGUSTO CERNI; WUELTON MARCELO MONTEIRO; UMBERTO ZOTTICH; MANUELA BERTO PUCCA. Anti-vaccination movements in the world and in Brazil. Revista da Sociedade Brasileira de Medicina Tropical, v. 55, 2022-05-20. (20/13176-3)

CARVALHO, ERICA S.; OLIVEIRA, ISADORA; NASCIMENTO, THAIS P.; NETO, ALEXANDRE VILHENA DA SILVA; LEAL, BRENDA A. S.; ARAUJO, FELIPE Q.; JULIAO, BRUNO F. V.; SOUZA, ANDREA R. N.; ABRAHIM, ANDREZA W.; MACEDO, BRUNA B. O.; et al. Prospecting Local Treatments Used in Conjunction with Antivenom Administration Following Envenomation Caused by Animals: A Systematic Review. TOXINS, v. 15, n. 5, p. 19-pg., 2023-04-28. (20/13176-3, 22/08964-8)

SALIMO, ZECA M.; BARROS, ANDRE L.; ADRIAO, ASENATE A. X.; RODRIGUES, ALINE M.; SARTIM, MARCO A.; DE OLIVEIRA, ISADORA S.; PUCCA, MANUELA B.; BAIA-DA-SILVA, DJANE C.; MONTEIRO, WUELTON M.; DE MELO, GISELY C.; et al. Toxins from Animal Venoms as a Potential Source of Antimalarials: A Comprehensive Review. TOXINS, v. 15, n. 6, p. 26-pg., 2023-06-01. (20/13176-3, 22/08964-8)

GALAN, LUIS E. B.; SILVA, VITORIA SOUZA; SILVA, VITORIA SANTOS; MONTE, ROMMEL C. C.; JATI, SEWBERT R. R.; OLIVEIRA, ISADORA S. S.; CERNI, FELIPE A. A.; MONTEIRO, WUELTON M. M.; SACHETT, JACQUELINE; DANTAS, DOMINGOS S. M.; et al. Acute mesenteric ischemia following lancehead snakebite: an unusual case report in the Northernmost Brazilian Amazon. FRONTIERS IN MEDICINE, v. 10, p. 7-pg., 2023-06-22. (20/13176-3, 22/08964-8)

CERNI, FELIPE; OLIVEIRA, ISADORA; CORDEIRO, FRANCIELLE; BORDON, KARLA; FERREIRA, ISABELA; MONTEIRO, WUELTON; ARANTES, ELIANE; CUNHA, THIAGO; PUCCA, MANUELA. The nociceptive response induced by different classes of Tityus serrulatus neurotoxins: The important role of Ts5 in venom-induced nociception. PLoS Neglected Tropical Diseases, v. 17, n. 1, p. 14-pg., 2023-01-01. (20/13176-3, 19/10173-6, 13/08216-2)

OLIVEIRA, ISADORA; COSTA, VICTOR; VERAS, FLAVIO; FERREIRA, ISABELA; CUNHA, FERNANDO; CUNHA, THIAGO; MONTEIRO, WUELTON; ARANTES, ELIANE; PUCCA, MANUELA. A Promising Biomolecule Able to Degrade Neutrophil Extracellular Traps: CdcPDE, a Rattlesnake Phosphodiesterase. TOXINS, v. 15, n. 1, p. 5-pg., 2023-01-01. (20/13176-3, 19/10173-6, 13/08216-2)

WIEZEL, GISELE A.; OLIVEIRA, ISADORA S.; FERREIRA, ISABELA G.; BORDON, KARLA C. F.; ARANTES, ELIANE C.. Hyperglycosylation impairs the inhibitory activity of rCdtPLI2, the first recombinant beta-phospholipase A₂ inhibitor. International Journal of Biological Macromolecules, v. 280, p. 12-pg., 2024-09-22. (21/11936-3, 09/54098-6, 19/10173-6, 20/13176-3, 17/00586-6)

WIEZEL, GISELE A.; OLIVEIRA, ISADORA S.; REIS, MOUZARLLEM B.; FERREIRA, ISABELA G.; CORDEIRO, KALYNKA R.; BORDON, KARLA C. F.; ARANTES, ELIANE C.. The complex repertoire of Tityus spp. venoms: Advances on their composition and pharmacological potential of their toxins. Biochimie, v. 220, p. 23-pg., 2024-01-11. (23/11264-0, 21/11936-3, 20/13176-3, 22/08964-8, 23/01083-9, 23/10400-8)

CAROLINA PETRI BERNARDES; ERNESTO LOPES PINHEIRO JUNIOR; ISABELA GOBBO FERREIRA; ISADORA SOUSA DE OLIVEIRA; NEIFE APARECIDA GUINAIM DOS SANTOS; SUELY VILELA SAMPAIO; ELIANE CANDIANI ARANTES; ANTONIO CARDOZO DOS SANTOS. Fraction of C. d. collilineatus venom containing crotapotin protects PC12 cells against MPP + toxicity by activating the NGF-signaling pathway. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 30, 2024-06-14. (19/10173-6, 11/23236-4, 20/13176-3)

BERNARDE, PAULO SERGIO; PUCCA, MANUELA BERTO; MOTA-DA-SILVA, AGEANE; DA FONSECA, WIRVEN LIMA; DE ALMEIDA, MARLLUS RAFAEL NEGREIROS; DE OLIVEIRA, ISADORA SOUSA; CERNI, FELIPE AUGUSTO; GOBBI GRAZZIOTIN, FELIPE; SARTIM, MARCO A.; SACHETT, JACQUELINE; et al. Bothrops bilineatus: An Arboreal Pitviper in the Amazon and Atlantic Forest. FRONTIERS IN IMMUNOLOGY, v. 12, DEC 15 2021. (20/13176-3, 16/50127-5)

OLIVEIRA, ISADORA; FERREIRA, ISABELA; JACOB, BEATRIZ; CARDENAS, KIARA; CERNI, FELIPE; BAIA-DA-SILVA, DJANE; ARANTES, ELIANE; MONTEIRO, WUELTON; PUCCA, MANUELA. Harnessing the Power of Venomous Animal-Derived Toxins against COVID-19. TOXINS, v. 15, n. 2, p. 12-pg., 2023-02-01. (20/13176-3, 21/11936-3, 19/10173-6)

OLIVEIRA, ISADORA S.; ANANIAS, CARLA B.; MEDEIROS, JILVANDO M.; FRANCO, MICHELLE V. S.; FERREIRA, ISABELA G.; CERNI, FELIPE A.; SANDRI, ELISEU A.; MONTEIRO, WUELTON M.; PUCCA, MANUELA B.. Medical Management after Lancehead Snakebite in North Amazon: A Case Report of Long-Term Disability. TOXINS, v. 14, n. 7, p. 9-pg., 2022-07-01. (20/13176-3)

DE OLIVEIRA, ISADORA SOUSA; PUCCA, MANUELA BERTO; FERREIRA, ISABELA GOBBO; CERNI, FELIPE AUGUSTO; JACOB, BEATRIZ DE CASSIA DA SILVA; WIEZEL, GISELE ADRIANO; PINHEIRO-JUNIOR, ERNESTO LOPES; CORDEIRO, FRANCIELLE ALMEIDA; FIGUEIREDO BORDON, KARLA DE CASTRO; ARANTES, ELIANE CANDIANI. State-of-the-art review of snake venom phosphodiesterases (svPDEs). Toxicon, v. 217, p. 10-pg., 2022-08-23. (17/00586-6, 20/13176-3)

ZOTTICH, UMBERTO; DE OLIVEIRA, ISADORA SOUSA; FEREIRA, ISABELA GOBBO; CERNI, FELIPE AUGUSTO; FIGUEIREDO, BORDON KARLA DE CASTRO; ARANTES, ELIANE CANDIANI; GOMES, VALDIRENE MOREIRA; DIAS, GERMANA BUENO; PUCCA, MANUELA BERTO. Antifungal activity of Rhopalurus crassicauda venom against Candida spp.. TOXICON-X, v. 14, p. 4-pg., 2022-06-01. (20/13176-3)

ALANO-DA-SILVA, NICOLY MALACHIZE; DE OLIVEIRA, ISADORA SOUSA; CARDOSO, IARA AIME; BORDON, KARLA DE CASTRO FIGUEIREDO; ARANTES, ELIANE CANDIANI. Exploring high molecular weight components in Tityus serrulatus venom. Toxicon, v. 255, p. 5-pg., 2025-01-17. (20/13176-3, 19/10173-6, 21/11547-7, 21/11936-3, 22/08964-8)

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)

WIEZEL, GISELE ADRIANO; OLIVEIRA, ISADORA SOUSA DE; ARANTES, ELIANE CANDIANI. Simplifying traditional approaches for accessible analysis of snake venom enzymes. Toxicon, v. 255, p. 5-pg., 2025-01-22. (20/13176-3, 17/00586-6, 21/11936-3, 19/10173-6)

REIS-FILHO, NELSON CARLOS; AGUIAR, THAYS KAROLYNE PONTE PRADO; RAMOS, DAFNIN LIMA DE SOUZA; MUELLER, BRUNA MARIAH MARTINS; DE OLIVEIRA, ISADORA SOUSA; CERNI, FELIPE AUGUSTO; PUCCA, MANUELA BERTO. Unprecedented report: First female monozygotic twins as carriers of Hutchinson-Gilford progeria syndrome. TRENDS IN PEDIATRICS, v. 5, n. 2, p. 5-pg., 2024-06-30. (20/13176-3)

DE MELLO, MARCELA M. BLASCKE; NEVES, VIVIANO GOMES DE OLIVEIRA; PARENTE, JULIANA MONTENEGRO; PERNOMIAN, LAENA; DE OLIVEIRA, ISADORA SOUSA; PEDERSOLI, CARINA AMARANTE; AWATA, WANESSA MAYUMI CARVALHO; TIRAPELLI, CARLOS RENATO; ARANTES, ELIANE CANDIANI; TOSTES, RITA DE CASSIA A.; et al. Sarcoplasmic reticulum calcium ATPase (SERCA) proteolysis by matrix metalloproteinase-2 contributes to vascular dysfunction in early hypertension. European Journal of Pharmacology, v. 983, p. 9-pg., 2024-09-07. (19/09174-8, 21/11936-3, 20/13176-3, 20/02619-1, 22/04888-5)

VAIYAPURI, SAKTHIVEL; KADAM, PRIYANKA; CHANDRASEKHARUNI, GNANESWAR; OLIVEIRA, ISADORA S.; SENTHILKUMARAN, SUBRAMANIAN; SALIM, ANIKA; PATEL, KETAN; SACHETT, JACQUELINE DE ALMEIDA GONCALVES; PUCCA, MANUELA B.. Multifaceted community health education programs as powerful tools to mitigate snakebite-induced deaths, disabilities, and socioeconomic burden. TOXICON-X, v. 17, p. 12-pg., 2022-12-28. (20/13176-3)

ISADORA SOUSA DE OLIVEIRA; NICOLY MALACHIZE ALANO-DA-SILVA; ISABELA GOBBO FERREIRA; FELIPE AUGUSTO CERNI; JACQUELINE DE ALMEIDA GONÇALVES SACHETT; WUELTON MARCELO MONTEIRO; MANUELA BERTO PUCCA; ELIANE CANDIANI ARANTES. Understanding the complexity of Tityus serrulatus venom: A focus on high molecular weight components. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 30, 2024-01-22. (22/08964-8, 21/11547-7, 19/10173-6, 20/13176-3, 21/11936-3)

GABRIELA DE OLIVEIRA ALMEIDA; ISADORA SOUSA DE OLIVEIRA; ELIANE CANDIANI ARANTES; SUELY VILELA SAMPAIO. Snake venom disintegrins update: insights about new findings. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 29, 2023-09-18. (20/15670-5, 22/08964-8, 22/04804-6, 19/10173-6, 22/05554-3, 20/13176-3, 21/11936-3)