Anti-Metalloprotease P-I Single-Domain Antibodies: Tools for Next-Generation Snakebite Antivenoms
Autor(a) principal: | |
---|---|
Data de Publicação: | 2022 |
Outros Autores: | , , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1155/2022/2748962 http://hdl.handle.net/11449/241445 |
Resumo: | In order to address the global antivenom crisis, novel antivenoms need to present high therapeutic efficacy, broad neutralization ability against systemic and local damage, sufficient safety, and cost-effectiveness. Due to biological characteristics of camelid single-domain antibodies (VHH) such as high affinity, their ability to penetrate dense tissues, and facility for genetic manipulation, their application in antivenoms has expanded considerably. VHHs that are active against the metalloprotease BjussuMP-II from the snake Bothrops jararacussu were selected. After isolation of BjussuMP-II, a camelid was immunized with the purified toxin in order to construct the recombinant phage library. Following a round of biopanning, 52% of the selected clones were able to recognize BjussuMP-II in an ELISA assay. After sequencing, seven sequence profiles were identified. One selected clone (VHH61) showed cross-reactivity to B. brazili venom, but did not recognize the Crotalus and Lachesis genera, indicating specificity for the Bothrops genus. Through in vitro tests, the capacity to neutralize the toxicity triggered by BjussuMP-II was observed. Circular dichroism spectroscopy indicated a robust secondary structure for VHH61, and the calculated melting temperature (TM) for the clone was 56.4°C. In silico analysis, through molecular docking of anti-BjussuMP-II VHHs with metalloprotease, revealed their potential interaction with amino acids present in regions critical for the toxin's conformation and stability. The findings suggest that anti-BjussuMP-II VHHs may be beneficial in the development of next-generation antivenoms. |
id |
UNSP_d4896d68d7b06695ff78800dc25af07d |
---|---|
oai_identifier_str |
oai:repositorio.unesp.br:11449/241445 |
network_acronym_str |
UNSP |
network_name_str |
Repositório Institucional da UNESP |
repository_id_str |
2946 |
spelling |
Anti-Metalloprotease P-I Single-Domain Antibodies: Tools for Next-Generation Snakebite AntivenomsIn order to address the global antivenom crisis, novel antivenoms need to present high therapeutic efficacy, broad neutralization ability against systemic and local damage, sufficient safety, and cost-effectiveness. Due to biological characteristics of camelid single-domain antibodies (VHH) such as high affinity, their ability to penetrate dense tissues, and facility for genetic manipulation, their application in antivenoms has expanded considerably. VHHs that are active against the metalloprotease BjussuMP-II from the snake Bothrops jararacussu were selected. After isolation of BjussuMP-II, a camelid was immunized with the purified toxin in order to construct the recombinant phage library. Following a round of biopanning, 52% of the selected clones were able to recognize BjussuMP-II in an ELISA assay. After sequencing, seven sequence profiles were identified. One selected clone (VHH61) showed cross-reactivity to B. brazili venom, but did not recognize the Crotalus and Lachesis genera, indicating specificity for the Bothrops genus. Through in vitro tests, the capacity to neutralize the toxicity triggered by BjussuMP-II was observed. Circular dichroism spectroscopy indicated a robust secondary structure for VHH61, and the calculated melting temperature (TM) for the clone was 56.4°C. In silico analysis, through molecular docking of anti-BjussuMP-II VHHs with metalloprotease, revealed their potential interaction with amino acids present in regions critical for the toxin's conformation and stability. The findings suggest that anti-BjussuMP-II VHHs may be beneficial in the development of next-generation antivenoms.Fundação Oswaldo Cruz Fiocruz Rondônia RondôniaCentro de Pesquisa em Medicina Tropical, RondôniaDepartamento de Biofísica e Farmacologia Instituto de Biociências UNESP BotucatuPlataforma Bi-Institucional Fiocruz-USP Ribeirão PretoInstituto Nacional de Ciência e Tecnologia de Epidemiologia da Amazônia Ocidental INCT-EpiAmOUniversidade Federal de Rondônia UNIR, RondôniaFundação Oswaldo Cruz Fiocruz Ceará EusébioDepartamento de Biofísica e Farmacologia Instituto de Biociências UNESP BotucatuRondôniaCentro de Pesquisa em Medicina TropicalUniversidade Estadual Paulista (UNESP)Universidade de São Paulo (USP)INCT-EpiAmOUNIREusébioSilva, Marcela C. S.Pereira, Soraya S.Gouveia, Marilia P.Luiz, Marcos B.Sousa, Rosa M. O.Kayano, Anderson M.Francisco, Aleff F. [UNESP]Prado, Nidiane D. R.Dill, Leandro S. M.Fontes, Marcos R. M. [UNESP]Zanchi, Fernando B.Stabeli, Rodrigo G.Soares, Andreimar M.Zuliani, Juliana P.Fernandes, Carla F. C.2023-03-01T21:03:34Z2023-03-01T21:03:34Z2022-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1155/2022/2748962BioMed Research International, v. 2022.2314-61412314-6133http://hdl.handle.net/11449/24144510.1155/2022/27489622-s2.0-85135248328Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBioMed Research Internationalinfo:eu-repo/semantics/openAccess2023-03-01T21:03:34Zoai:repositorio.unesp.br:11449/241445Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-06T00:14:23.838285Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Anti-Metalloprotease P-I Single-Domain Antibodies: Tools for Next-Generation Snakebite Antivenoms |
title |
Anti-Metalloprotease P-I Single-Domain Antibodies: Tools for Next-Generation Snakebite Antivenoms |
spellingShingle |
Anti-Metalloprotease P-I Single-Domain Antibodies: Tools for Next-Generation Snakebite Antivenoms Silva, Marcela C. S. |
title_short |
Anti-Metalloprotease P-I Single-Domain Antibodies: Tools for Next-Generation Snakebite Antivenoms |
title_full |
Anti-Metalloprotease P-I Single-Domain Antibodies: Tools for Next-Generation Snakebite Antivenoms |
title_fullStr |
Anti-Metalloprotease P-I Single-Domain Antibodies: Tools for Next-Generation Snakebite Antivenoms |
title_full_unstemmed |
Anti-Metalloprotease P-I Single-Domain Antibodies: Tools for Next-Generation Snakebite Antivenoms |
title_sort |
Anti-Metalloprotease P-I Single-Domain Antibodies: Tools for Next-Generation Snakebite Antivenoms |
author |
Silva, Marcela C. S. |
author_facet |
Silva, Marcela C. S. Pereira, Soraya S. Gouveia, Marilia P. Luiz, Marcos B. Sousa, Rosa M. O. Kayano, Anderson M. Francisco, Aleff F. [UNESP] Prado, Nidiane D. R. Dill, Leandro S. M. Fontes, Marcos R. M. [UNESP] Zanchi, Fernando B. Stabeli, Rodrigo G. Soares, Andreimar M. Zuliani, Juliana P. Fernandes, Carla F. C. |
author_role |
author |
author2 |
Pereira, Soraya S. Gouveia, Marilia P. Luiz, Marcos B. Sousa, Rosa M. O. Kayano, Anderson M. Francisco, Aleff F. [UNESP] Prado, Nidiane D. R. Dill, Leandro S. M. Fontes, Marcos R. M. [UNESP] Zanchi, Fernando B. Stabeli, Rodrigo G. Soares, Andreimar M. Zuliani, Juliana P. Fernandes, Carla F. C. |
author2_role |
author author author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Rondônia Centro de Pesquisa em Medicina Tropical Universidade Estadual Paulista (UNESP) Universidade de São Paulo (USP) INCT-EpiAmO UNIR Eusébio |
dc.contributor.author.fl_str_mv |
Silva, Marcela C. S. Pereira, Soraya S. Gouveia, Marilia P. Luiz, Marcos B. Sousa, Rosa M. O. Kayano, Anderson M. Francisco, Aleff F. [UNESP] Prado, Nidiane D. R. Dill, Leandro S. M. Fontes, Marcos R. M. [UNESP] Zanchi, Fernando B. Stabeli, Rodrigo G. Soares, Andreimar M. Zuliani, Juliana P. Fernandes, Carla F. C. |
description |
In order to address the global antivenom crisis, novel antivenoms need to present high therapeutic efficacy, broad neutralization ability against systemic and local damage, sufficient safety, and cost-effectiveness. Due to biological characteristics of camelid single-domain antibodies (VHH) such as high affinity, their ability to penetrate dense tissues, and facility for genetic manipulation, their application in antivenoms has expanded considerably. VHHs that are active against the metalloprotease BjussuMP-II from the snake Bothrops jararacussu were selected. After isolation of BjussuMP-II, a camelid was immunized with the purified toxin in order to construct the recombinant phage library. Following a round of biopanning, 52% of the selected clones were able to recognize BjussuMP-II in an ELISA assay. After sequencing, seven sequence profiles were identified. One selected clone (VHH61) showed cross-reactivity to B. brazili venom, but did not recognize the Crotalus and Lachesis genera, indicating specificity for the Bothrops genus. Through in vitro tests, the capacity to neutralize the toxicity triggered by BjussuMP-II was observed. Circular dichroism spectroscopy indicated a robust secondary structure for VHH61, and the calculated melting temperature (TM) for the clone was 56.4°C. In silico analysis, through molecular docking of anti-BjussuMP-II VHHs with metalloprotease, revealed their potential interaction with amino acids present in regions critical for the toxin's conformation and stability. The findings suggest that anti-BjussuMP-II VHHs may be beneficial in the development of next-generation antivenoms. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-01-01 2023-03-01T21:03:34Z 2023-03-01T21:03:34Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1155/2022/2748962 BioMed Research International, v. 2022. 2314-6141 2314-6133 http://hdl.handle.net/11449/241445 10.1155/2022/2748962 2-s2.0-85135248328 |
url |
http://dx.doi.org/10.1155/2022/2748962 http://hdl.handle.net/11449/241445 |
identifier_str_mv |
BioMed Research International, v. 2022. 2314-6141 2314-6133 10.1155/2022/2748962 2-s2.0-85135248328 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
BioMed Research International |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808129599072108544 |