Interaction between Shiga Toxin and Monoclonal Antibodies: Binding Characteristics and in Vitro Neutralizing Abilities
Autor(a) principal: | |
---|---|
Data de Publicação: | 2012 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/35200 http://dx.doi.org/10.3390/toxins4090729 |
Resumo: | Monoclonal antibodies (MAbs) have been employed either for diagnosis or treatment of infections caused by different pathogens. Specifically for Shiga toxin-producing Escherichia coli (STEC), numerous immunoassays have been developed for STEC diagnosis, showing variability in sensitivity and specificity when evaluated by reference laboratories, and no therapy or vaccines are currently approved. Thus, the aim of this work was the characterization of the interaction between MAbs against Stx1 and Stx2 toxins and their neutralizing abilities to enable their use as tools for diagnosis and therapy. the selected clones designated 3E2 (anti-Stx1) and 2E11 (anti-Stx2) were classified as IgG1. 3E2 recognized the B subunit of Stx1 with an affinity constant of 2.5 x 10(-10) M, detected as little as 6.2 ng of Stx1 and was stable up to 50 degrees C. in contrast, 2E11 recognized the A subunit of Stx2, was stable up to 70 degrees C, had a high dissociation constant of 6.1 x 10(-10) M, and detected as little as 12.5 ng of Stx2. Neutralization tests showed that 160 ng of 3E2 MAb inhibited 80% of Stx1 activity and 500 mu g 2E11 MAb were required for 60% inhibition of Stx2 activity. These MAb amounts reversed 25 to 80% of the cytotoxicity triggered by different STEC isolates. in conclusion, these MAbs show suitable characteristics for their use in STEC diagnosis and encourage future studies to investigate their protective efficacy. |
id |
UFSP_07583c0a022022622276479301a86a3a |
---|---|
oai_identifier_str |
oai:repositorio.unifesp.br:11600/35200 |
network_acronym_str |
UFSP |
network_name_str |
Repositório Institucional da UNIFESP |
repository_id_str |
3465 |
spelling |
Rocha, Leticia B.Luz, Daniela E.Moraes, Claudia T. P.Caravelli, AndressaFernandes, IreneGuth, Beatriz Ernestina Cabilio [UNIFESP]Horton, Denise S. P. Q.Piazza, Roxane M. F.Butantan InstUniversidade Federal de São Paulo (UNIFESP)2016-01-24T14:27:36Z2016-01-24T14:27:36Z2012-09-01Toxins. Basel: Mdpi Ag, v. 4, n. 9, p. 729-747, 2012.2072-6651http://repositorio.unifesp.br/handle/11600/35200http://dx.doi.org/10.3390/toxins4090729WOS000315405900007.pdf10.3390/toxins4090729WOS:000315405900007Monoclonal antibodies (MAbs) have been employed either for diagnosis or treatment of infections caused by different pathogens. Specifically for Shiga toxin-producing Escherichia coli (STEC), numerous immunoassays have been developed for STEC diagnosis, showing variability in sensitivity and specificity when evaluated by reference laboratories, and no therapy or vaccines are currently approved. Thus, the aim of this work was the characterization of the interaction between MAbs against Stx1 and Stx2 toxins and their neutralizing abilities to enable their use as tools for diagnosis and therapy. the selected clones designated 3E2 (anti-Stx1) and 2E11 (anti-Stx2) were classified as IgG1. 3E2 recognized the B subunit of Stx1 with an affinity constant of 2.5 x 10(-10) M, detected as little as 6.2 ng of Stx1 and was stable up to 50 degrees C. in contrast, 2E11 recognized the A subunit of Stx2, was stable up to 70 degrees C, had a high dissociation constant of 6.1 x 10(-10) M, and detected as little as 12.5 ng of Stx2. Neutralization tests showed that 160 ng of 3E2 MAb inhibited 80% of Stx1 activity and 500 mu g 2E11 MAb were required for 60% inhibition of Stx2 activity. These MAb amounts reversed 25 to 80% of the cytotoxicity triggered by different STEC isolates. in conclusion, these MAbs show suitable characteristics for their use in STEC diagnosis and encourage future studies to investigate their protective efficacy.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Butantan Inst, Bacteriol Lab, São Paulo, BrazilButantan Inst, Immunopathol Lab, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol, Escola Paulista Med, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol, Escola Paulista Med, São Paulo, BrazilWeb of Science729-747engMdpi AgToxinsStx1Stx2monoclonal antibodiesbindingstabilitydetectionneutralizing abilityspecificityInteraction between Shiga Toxin and Monoclonal Antibodies: Binding Characteristics and in Vitro Neutralizing Abilitiesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALWOS000315405900007.pdfapplication/pdf1412043${dspace.ui.url}/bitstream/11600/35200/1/WOS000315405900007.pdf6e57a67bf8db649c0ce35f1778303f15MD51open accessTEXTWOS000315405900007.pdf.txtWOS000315405900007.pdf.txtExtracted texttext/plain51153${dspace.ui.url}/bitstream/11600/35200/2/WOS000315405900007.pdf.txtcb36f8b66705aa08e421d32c6737a682MD52open access11600/352002023-02-15 09:10:18.174open accessoai:repositorio.unifesp.br:11600/35200Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-05-25T12:30:36.470250Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Interaction between Shiga Toxin and Monoclonal Antibodies: Binding Characteristics and in Vitro Neutralizing Abilities |
title |
Interaction between Shiga Toxin and Monoclonal Antibodies: Binding Characteristics and in Vitro Neutralizing Abilities |
spellingShingle |
Interaction between Shiga Toxin and Monoclonal Antibodies: Binding Characteristics and in Vitro Neutralizing Abilities Rocha, Leticia B. Stx1 Stx2 monoclonal antibodies binding stability detection neutralizing ability specificity |
title_short |
Interaction between Shiga Toxin and Monoclonal Antibodies: Binding Characteristics and in Vitro Neutralizing Abilities |
title_full |
Interaction between Shiga Toxin and Monoclonal Antibodies: Binding Characteristics and in Vitro Neutralizing Abilities |
title_fullStr |
Interaction between Shiga Toxin and Monoclonal Antibodies: Binding Characteristics and in Vitro Neutralizing Abilities |
title_full_unstemmed |
Interaction between Shiga Toxin and Monoclonal Antibodies: Binding Characteristics and in Vitro Neutralizing Abilities |
title_sort |
Interaction between Shiga Toxin and Monoclonal Antibodies: Binding Characteristics and in Vitro Neutralizing Abilities |
author |
Rocha, Leticia B. |
author_facet |
Rocha, Leticia B. Luz, Daniela E. Moraes, Claudia T. P. Caravelli, Andressa Fernandes, Irene Guth, Beatriz Ernestina Cabilio [UNIFESP] Horton, Denise S. P. Q. Piazza, Roxane M. F. |
author_role |
author |
author2 |
Luz, Daniela E. Moraes, Claudia T. P. Caravelli, Andressa Fernandes, Irene Guth, Beatriz Ernestina Cabilio [UNIFESP] Horton, Denise S. P. Q. Piazza, Roxane M. F. |
author2_role |
author author author author author author author |
dc.contributor.institution.none.fl_str_mv |
Butantan Inst Universidade Federal de São Paulo (UNIFESP) |
dc.contributor.author.fl_str_mv |
Rocha, Leticia B. Luz, Daniela E. Moraes, Claudia T. P. Caravelli, Andressa Fernandes, Irene Guth, Beatriz Ernestina Cabilio [UNIFESP] Horton, Denise S. P. Q. Piazza, Roxane M. F. |
dc.subject.eng.fl_str_mv |
Stx1 Stx2 monoclonal antibodies binding stability detection neutralizing ability specificity |
topic |
Stx1 Stx2 monoclonal antibodies binding stability detection neutralizing ability specificity |
description |
Monoclonal antibodies (MAbs) have been employed either for diagnosis or treatment of infections caused by different pathogens. Specifically for Shiga toxin-producing Escherichia coli (STEC), numerous immunoassays have been developed for STEC diagnosis, showing variability in sensitivity and specificity when evaluated by reference laboratories, and no therapy or vaccines are currently approved. Thus, the aim of this work was the characterization of the interaction between MAbs against Stx1 and Stx2 toxins and their neutralizing abilities to enable their use as tools for diagnosis and therapy. the selected clones designated 3E2 (anti-Stx1) and 2E11 (anti-Stx2) were classified as IgG1. 3E2 recognized the B subunit of Stx1 with an affinity constant of 2.5 x 10(-10) M, detected as little as 6.2 ng of Stx1 and was stable up to 50 degrees C. in contrast, 2E11 recognized the A subunit of Stx2, was stable up to 70 degrees C, had a high dissociation constant of 6.1 x 10(-10) M, and detected as little as 12.5 ng of Stx2. Neutralization tests showed that 160 ng of 3E2 MAb inhibited 80% of Stx1 activity and 500 mu g 2E11 MAb were required for 60% inhibition of Stx2 activity. These MAb amounts reversed 25 to 80% of the cytotoxicity triggered by different STEC isolates. in conclusion, these MAbs show suitable characteristics for their use in STEC diagnosis and encourage future studies to investigate their protective efficacy. |
publishDate |
2012 |
dc.date.issued.fl_str_mv |
2012-09-01 |
dc.date.accessioned.fl_str_mv |
2016-01-24T14:27:36Z |
dc.date.available.fl_str_mv |
2016-01-24T14:27:36Z |
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 |
Toxins. Basel: Mdpi Ag, v. 4, n. 9, p. 729-747, 2012. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/35200 http://dx.doi.org/10.3390/toxins4090729 |
dc.identifier.issn.none.fl_str_mv |
2072-6651 |
dc.identifier.file.none.fl_str_mv |
WOS000315405900007.pdf |
dc.identifier.doi.none.fl_str_mv |
10.3390/toxins4090729 |
dc.identifier.wos.none.fl_str_mv |
WOS:000315405900007 |
identifier_str_mv |
Toxins. Basel: Mdpi Ag, v. 4, n. 9, p. 729-747, 2012. 2072-6651 WOS000315405900007.pdf 10.3390/toxins4090729 WOS:000315405900007 |
url |
http://repositorio.unifesp.br/handle/11600/35200 http://dx.doi.org/10.3390/toxins4090729 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Toxins |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
729-747 |
dc.publisher.none.fl_str_mv |
Mdpi Ag |
publisher.none.fl_str_mv |
Mdpi Ag |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
instname_str |
Universidade Federal de São Paulo (UNIFESP) |
instacron_str |
UNIFESP |
institution |
UNIFESP |
reponame_str |
Repositório Institucional da UNIFESP |
collection |
Repositório Institucional da UNIFESP |
bitstream.url.fl_str_mv |
${dspace.ui.url}/bitstream/11600/35200/1/WOS000315405900007.pdf ${dspace.ui.url}/bitstream/11600/35200/2/WOS000315405900007.pdf.txt |
bitstream.checksum.fl_str_mv |
6e57a67bf8db649c0ce35f1778303f15 cb36f8b66705aa08e421d32c6737a682 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
repository.mail.fl_str_mv |
|
_version_ |
1783460300578619392 |