Camelid single-domain antibodies (VHHs) against crotoxin: a basis for developing modular building blocks for the enhancement of treatment or diagnosis of crotalic envenoming

Detalhes bibliográficos
Autor(a) principal: Luiz, Marcos B.
Data de Publicação: 2018
Outros Autores: Pereira, Soraya S., Prado, Nidiane D. R., Gonçalves, Naan R., Kayano, Anderson M., Moreira-Dill, Leandro S., Sobrinho, Juliana C., Zanchi, Fernando B., Fuly, André L., Fernandes, Cleberson F., Zuliani, Juliana P., Soares, Andreimar M., Stabeli, Rodrigo G., Fernandes, Carla F. C.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/35522
Resumo: 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 throughWestern 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.
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spelling Camelid single-domain antibodies (VHHs) against crotoxin: a basis for developing modular building blocks for the enhancement of treatment or diagnosis of crotalic envenomingCrotoxinaCrotoxinCrotalus cascavellaAbstract: 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 throughWestern 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.Toxins2018-09-05T18:42:08Z2018-09-05T18:42:08Z2018-03info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfLUIZ, M. B. 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. Toxins, v. 10, n. 142, p. 1-20, mar. 2018.2072-6651http://www.repositorio.ufc.br/handle/riufc/35522Luiz, Marcos B.Pereira, Soraya S.Prado, Nidiane D. R.Gonçalves, Naan R.Kayano, Anderson M.Moreira-Dill, Leandro S.Sobrinho, Juliana C.Zanchi, Fernando B.Fuly, André L.Fernandes, Cleberson F.Zuliani, Juliana P.Soares, Andreimar M.Stabeli, Rodrigo G.Fernandes, Carla F. C.engreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2019-01-25T13:52:17Zoai:repositorio.ufc.br:riufc/35522Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:19:59.350122Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Camelid single-domain antibodies (VHHs) against crotoxin: a basis for developing modular building blocks for the enhancement of treatment or diagnosis of crotalic envenoming
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
spellingShingle Camelid single-domain antibodies (VHHs) against crotoxin: a basis for developing modular building blocks for the enhancement of treatment or diagnosis of crotalic envenoming
Luiz, Marcos B.
Crotoxina
Crotoxin
Crotalus cascavella
title_short Camelid single-domain antibodies (VHHs) against crotoxin: a basis for developing modular building blocks for the enhancement of treatment or diagnosis of crotalic envenoming
title_full Camelid single-domain antibodies (VHHs) against crotoxin: a basis for developing modular building blocks for the enhancement of treatment or diagnosis of crotalic envenoming
title_fullStr Camelid single-domain antibodies (VHHs) against crotoxin: a basis for developing modular building blocks for the enhancement of treatment or diagnosis of crotalic envenoming
title_full_unstemmed Camelid single-domain antibodies (VHHs) against crotoxin: a basis for developing modular building blocks for the enhancement of treatment or diagnosis of crotalic envenoming
title_sort Camelid single-domain antibodies (VHHs) against crotoxin: a basis for developing modular building blocks for the enhancement of treatment or diagnosis of crotalic envenoming
author Luiz, Marcos B.
author_facet Luiz, Marcos B.
Pereira, Soraya S.
Prado, Nidiane D. R.
Gonçalves, Naan R.
Kayano, Anderson M.
Moreira-Dill, Leandro S.
Sobrinho, Juliana C.
Zanchi, Fernando B.
Fuly, André L.
Fernandes, Cleberson F.
Zuliani, Juliana P.
Soares, Andreimar M.
Stabeli, Rodrigo G.
Fernandes, Carla F. C.
author_role author
author2 Pereira, Soraya S.
Prado, Nidiane D. R.
Gonçalves, Naan R.
Kayano, Anderson M.
Moreira-Dill, Leandro S.
Sobrinho, Juliana C.
Zanchi, Fernando B.
Fuly, André L.
Fernandes, Cleberson F.
Zuliani, Juliana P.
Soares, Andreimar M.
Stabeli, Rodrigo G.
Fernandes, Carla F. C.
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Luiz, Marcos B.
Pereira, Soraya S.
Prado, Nidiane D. R.
Gonçalves, Naan R.
Kayano, Anderson M.
Moreira-Dill, Leandro S.
Sobrinho, Juliana C.
Zanchi, Fernando B.
Fuly, André L.
Fernandes, Cleberson F.
Zuliani, Juliana P.
Soares, Andreimar M.
Stabeli, Rodrigo G.
Fernandes, Carla F. C.
dc.subject.por.fl_str_mv Crotoxina
Crotoxin
Crotalus cascavella
topic Crotoxina
Crotoxin
Crotalus cascavella
description 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 throughWestern 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.
publishDate 2018
dc.date.none.fl_str_mv 2018-09-05T18:42:08Z
2018-09-05T18:42:08Z
2018-03
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 LUIZ, M. B. 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. Toxins, v. 10, n. 142, p. 1-20, mar. 2018.
2072-6651
http://www.repositorio.ufc.br/handle/riufc/35522
identifier_str_mv LUIZ, M. B. 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. Toxins, v. 10, n. 142, p. 1-20, mar. 2018.
2072-6651
url http://www.repositorio.ufc.br/handle/riufc/35522
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Toxins
publisher.none.fl_str_mv Toxins
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Ceará (UFC)
instname:Universidade Federal do Ceará (UFC)
instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
instacron_str UFC
institution UFC
reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
collection Repositório Institucional da Universidade Federal do Ceará (UFC)
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
repository.mail.fl_str_mv bu@ufc.br || repositorio@ufc.br
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