Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay

Bibliographic Details
Main Author: De-Simone, Salvatore Giovanni
Publication Date: 2021
Other Authors: Gomes, Larissa Rodrigues, Napoleão-Pego, Paloma, Lechuga, Guilherme Curty, de Pina, Jorge Soares, Silva, Flávio Rocha da
Format: Article
Language: eng
Source: Repositório Institucional da FIOCRUZ (ARCA)
DOI: 10.3390/VACCINES9040313
Download full: https://arca.fiocruz.br/handle/icict/59627
Summary: Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças de Populações Negligenciadas Rio de Janeiro, RJ, Brasil.
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spelling De-Simone, Salvatore GiovanniGomes, Larissa RodriguesNapoleão-Pego, PalomaLechuga, Guilherme Curtyde Pina, Jorge SoaresSilva, Flávio Rocha da2023-07-18T03:00:07Z2023-07-18T03:00:07Z2021DE-SIMONE, Salvatore Giovanni etal. Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay. Vaccines, Switzerland, v. 9, n. 4, p. 1-14, 2021.2076-393Xhttps://arca.fiocruz.br/handle/icict/5962710.3390/VACCINES90403132076-393XengMDPIEpitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assayinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças de Populações Negligenciadas Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças de Populações Negligenciadas Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças de Populações Negligenciadas Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças de Populações Negligenciadas Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças de Populações Negligenciadas Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças de Populações Negligenciadas Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças de Populações Negligenciadas Rio de Janeiro, RJ, Brasil.Background: The diphtheria toxoid antigen is a major component in pediatric and booster combination vaccines and is known to raise a protective humoral immune response upon vaccination. Although antibodies are considered critical for diphtheria protection, little is known about the antigenic determinants that maintain humoral immunity. Methods: One-hundred and twelve 15 mer peptides covering the entire sequence of diphtheria toxin (DTx) protein were prepared by SPOT synthesis. The immunoreactivity of membrane-bound peptides with sera from mice immunized with a triple DTP vaccine allowed mapping of continuous B-cell epitopes, topological studies, multiantigen peptide (MAP) synthesis, and Enzyme-Linked Immunosorbent Assay (ELISA) development. Results: Twenty epitopes were identified, with two being in the signal peptide, five in the catalytic domain (CD), seven in the HBFT domain, and five in the receptor-binding domain (RBD). Two 17 mer (CB/Tx-2/12 and CB/DTx-4-13) derived biepitope peptides linked by a Gly-Gly spacer were chemically synthesized. The peptides were used as antigens to coat ELISA plates and assayed with human (huVS) and mice vaccinated sera (miVS) for in vitro diagnosis of diphtheria. The assay proved to be highly sensitive (99.96%) and specific (100%) for huVS and miVS and, when compared with a commercial ELISA test, demonstrated a high performance. Conclusions: Our work displayed the complete picture of the linear B cell IgG response epitope of the DTx responsible for the protective effect and demonstrated sufficient specificity and eligibility for phase IIB studies of some epitopes to develop new and fast diagnostic assays.Diphtheria toxinB epitopesEpitope mappingbi-specific peptidesELISADiagnostic performanceinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txttext/plain1748https://arca.fiocruz.br/bitstreams/64278e68-968e-42de-a778-4f0d82045075/download8a4605be74aa9ea9d79846c1fba20a33MD51falseAnonymousREADORIGINAL10.3390_vaccines9040313.pdfapplication/pdf2424544https://arca.fiocruz.br/bitstreams/01d88bdd-17e4-43b6-b888-6f06054fabed/download0bf43d6f754270c0a8ed355e05faffb5MD52trueAnonymousREADTEXT10.3390_vaccines9040313.pdf.txt10.3390_vaccines9040313.pdf.txtExtracted texttext/plain68747https://arca.fiocruz.br/bitstreams/0268409a-d285-40bf-9c1b-1553c8b07811/download835fc1228d472bc09f37b70b4f200e45MD55falseAnonymousREADTHUMBNAIL10.3390_vaccines9040313.pdf.jpg10.3390_vaccines9040313.pdf.jpgGenerated Thumbnailimage/jpeg5656https://arca.fiocruz.br/bitstreams/7d4e801a-fd48-4781-b04f-9dd1f1253eed/download9f2712061247e68a5350dd188dc1543eMD56falseAnonymousREADicict/596272025-07-30 02:30:04.41open.accessoai:arca.fiocruz.br:icict/59627https://arca.fiocruz.brRepositório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352025-07-30T05:30:04Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)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
dc.title.none.fl_str_mv Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay
title Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay
spellingShingle Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay
De-Simone, Salvatore Giovanni
Diphtheria toxin
B epitopes
Epitope mapping
bi-specific peptides
ELISA
Diagnostic performance
title_short Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay
title_full Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay
title_fullStr Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay
title_full_unstemmed Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay
title_sort Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay
author De-Simone, Salvatore Giovanni
author_facet De-Simone, Salvatore Giovanni
Gomes, Larissa Rodrigues
Napoleão-Pego, Paloma
Lechuga, Guilherme Curty
de Pina, Jorge Soares
Silva, Flávio Rocha da
author_role author
author2 Gomes, Larissa Rodrigues
Napoleão-Pego, Paloma
Lechuga, Guilherme Curty
de Pina, Jorge Soares
Silva, Flávio Rocha da
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv De-Simone, Salvatore Giovanni
Gomes, Larissa Rodrigues
Napoleão-Pego, Paloma
Lechuga, Guilherme Curty
de Pina, Jorge Soares
Silva, Flávio Rocha da
dc.subject.en.none.fl_str_mv Diphtheria toxin
B epitopes
Epitope mapping
bi-specific peptides
ELISA
Diagnostic performance
topic Diphtheria toxin
B epitopes
Epitope mapping
bi-specific peptides
ELISA
Diagnostic performance
description Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças de Populações Negligenciadas Rio de Janeiro, RJ, Brasil.
publishDate 2021
dc.date.issued.fl_str_mv 2021
dc.date.accessioned.fl_str_mv 2023-07-18T03:00:07Z
dc.date.available.fl_str_mv 2023-07-18T03:00:07Z
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.citation.fl_str_mv DE-SIMONE, Salvatore Giovanni etal. Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay. Vaccines, Switzerland, v. 9, n. 4, p. 1-14, 2021.
dc.identifier.uri.fl_str_mv https://arca.fiocruz.br/handle/icict/59627
dc.identifier.issn.none.fl_str_mv 2076-393X
dc.identifier.doi.none.fl_str_mv 10.3390/VACCINES9040313
dc.identifier.eissn.none.fl_str_mv 2076-393X
identifier_str_mv DE-SIMONE, Salvatore Giovanni etal. Epitope mapping of the diphtheria toxin and development of an ELISA-specific diagnostic assay. Vaccines, Switzerland, v. 9, n. 4, p. 1-14, 2021.
2076-393X
10.3390/VACCINES9040313
url https://arca.fiocruz.br/handle/icict/59627
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.publisher.none.fl_str_mv MDPI
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dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
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