| Author | Luiz, Marcos Barros | |
| Author | Pereira, Soraya Santos | |
| Author | Prado, Nidiane Dantas Reis | |
| Author | Gonçalves, Naan Rodrigues | |
| Author | Kayano, Anderson Makoto | |
| Author | Moreira-Dill, Leandro Soares | |
| Author | Sobrinho, Juliana Conceição | |
| Author | Zanchi, Fernando Berton | |
| Author | Fuly, André Lopes | |
| Author | Fernandes, Cleberson Freitas | |
| Author | Zuliani, Juliana Pavan Zuliani | |
| Author | Soares, Andreimar Martins | |
| Author | Stabeli, Rodrigo Guerino | |
| Author | Fernandes, Carla Freire Celedonio | |
| Access date | 2024-12-18T13:09:45Z | |
| Available date | 2024-12-18T13:09:45Z | |
| Document date | 2018 | |
| Citation | Luiz, Marcos Barros; Pereira, Soraya Santos; Prado, Nidiane Dantas Reis. et al. Camelid Single-Domain Antibodies (VHHs) against Crotoxin: A Basis for Developing Modular Building Blocks for the Enhancement of Treatment or Diagnosis of Crotalic Envenoming. Porto Velho, Brasil. Toxins Journal, v. 10(4), n. 142, 2018. | en_US |
| ISSN | 2072-6651 | en_US |
| URI | https://www.arca.fiocruz.br/handle/icict/67668 | |
| Sponsorship | FINEP
CAPES | en_US |
| Language | eng | en_US |
| Publisher | Toxins | en_US |
| Rights | open access | en_US |
| Title | Camelid Single-Domain Antibodies (VHHs) against Crotoxin: A Basis for Developing Modular Building Blocks for the Enhancement of Treatment or Diagnosis of Crotalic Envenoming | en_US |
| Type | Article | en_US |
| DOI | 10.3390/toxins10040142 | |
| Abstract | Toxic effects triggered by crotalic envenoming are mainly related to crotoxin (CTX), composed of a phospholipase A2 (CB) and a subunit with no toxic activity (CA). Camelids produce immunoglobulins G devoid of light chains, in which the antigen recognition domain is called VHH. Given their unique characteristics, VHHs were selected using Phage Display against CTX from Crotalus durissus terrificus. After three rounds of biopanning, four sequence profiles for CB (KF498602, KF498603, KF498604, and KF498605) and one for CA (KF498606) were revealed. All clones presented the VHH hallmark in FR2 and a long CDR3, with the exception of KF498606. After expressing pET22b-VHHs in E. coli, approximately 2 to 6 mg of protein per liter of culture were obtained. When tested for cross-reactivity, VHHs presented specificity for the Crotalus genus and were capable of recognizing CB through Western blot. KF498602 and KF498604 showed thermostability, and displayed affinity constants for CTX in the micro or nanomolar range. They inhibited in vitro CTX PLA2 activity, and CB cytotoxicity. Furthermore, KF498604 inhibited the CTX-induced myotoxicity in mice by 78.8%. Molecular docking revealed that KF498604 interacts with the CA–CB interface of CTX, seeming to block substrate access. Selected VHHs may be alternatives for the crotalic envenoming treatment. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brasil. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz Rondônia. Porto Velho, RO, Brasil / Fundação Universidade Federal de Rondônia. Porto Velho, RO, Brasil. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Fiocruz USP. Plataforma Bi-institucional de Pesquisa em Medicina Translacional. Ribeirão Preto, SP, Brasil. | en_US |
| Affilliation | Instituto Federal de Educação, Ciência e Tecnologia de Rondônia, IFRO. Guajará-Mirim, RO, Brasil. | en_US |
| Affilliation | Universidade Federal Fluminense, UFF. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Embrapa Agroindústria Tropical. Fortaleza, CE, Brasil. | en_US |
| Affilliation | Centro Universitário São Lucas, UniSL. Porto Velho, RO, Brasil. | en_US |
| Affilliation | Departamento de Medicina da Universidade Federal de São Carlos, Demed-UFSCAR. São Carlos, SP, Brasil. | en_US |
| Subject | crotoxin | en_US |
| Subject | CB | en_US |
| Subject | VHH | en_US |
| Subject | Crotalus durissus terrificus | en_US |
| DeCS | Crotoxin | en_US |
| DeCS | Antibodies VHH | en_US |
| DeCS | Crotalus cascavella | en_US |
