Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | The Journal of venomous animals and toxins including tropical diseases (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992021000100310 |
Resumo: | Abstract Background Tetanus toxin blocks the release of the inhibitory neurotransmitters in the central nervous system and causes tetanus and its main form of prevention is through vaccination. The vaccine is produced by inactivation of tetanus toxin with formaldehyde, which may cause side effects. An alternative way is the use of ionizing radiation for inactivation of the toxin and also to improve the potential immunogenic response and to reduce the post-vaccination side effects. Therefore, the aim of this study was to characterize the tetanus toxin structure after different doses of ionizing radiation of 60Co. Methods Irradiated and native tetanus toxin was characterized by SDS PAGE in reducing and non-reducing conditions and MALD-TOF. Enzymatic activity was measured by FRET substrate. Also, antigenic properties were assessed by ELISA and Western Blot data. Results Characterization analysis revealed gradual modification on the tetanus toxin structure according to doses increase. Also, fragmentation and possible aggregations of the protein fragments were observed in higher doses. In the analysis of peptide preservation by enzymatic digestion and mass spectrometry, there was a slight modification in the identification up to the dose of 4 kGy. At subsequent doses, peptide identification was minimal. The analysis of the enzymatic activity by fluorescence showed 35 % attenuation in the activity even at higher doses. In the antigenic evaluation, anti-tetanus toxin antibodies were detected against the irradiated toxins at the different doses, with a gradual decrease as the dose increased, but remaining at satisfactory levels. Conclusion Ionizing radiation promoted structural changes in the tetanus toxin such as fragmentation and/or aggregation and attenuation of enzymatic activity as the dose increased, but antigenic recognition of the toxin remained at good levels indicating its possible use as an immunogen. However, studies of enzymatic activity of tetanus toxin irradiated with doses above 8 kGy should be further analyzed. |
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The Journal of venomous animals and toxins including tropical diseases (Online) |
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Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiationRadiationFragmentationEnzymatic activityGamma raysAbstract Background Tetanus toxin blocks the release of the inhibitory neurotransmitters in the central nervous system and causes tetanus and its main form of prevention is through vaccination. The vaccine is produced by inactivation of tetanus toxin with formaldehyde, which may cause side effects. An alternative way is the use of ionizing radiation for inactivation of the toxin and also to improve the potential immunogenic response and to reduce the post-vaccination side effects. Therefore, the aim of this study was to characterize the tetanus toxin structure after different doses of ionizing radiation of 60Co. Methods Irradiated and native tetanus toxin was characterized by SDS PAGE in reducing and non-reducing conditions and MALD-TOF. Enzymatic activity was measured by FRET substrate. Also, antigenic properties were assessed by ELISA and Western Blot data. Results Characterization analysis revealed gradual modification on the tetanus toxin structure according to doses increase. Also, fragmentation and possible aggregations of the protein fragments were observed in higher doses. In the analysis of peptide preservation by enzymatic digestion and mass spectrometry, there was a slight modification in the identification up to the dose of 4 kGy. At subsequent doses, peptide identification was minimal. The analysis of the enzymatic activity by fluorescence showed 35 % attenuation in the activity even at higher doses. In the antigenic evaluation, anti-tetanus toxin antibodies were detected against the irradiated toxins at the different doses, with a gradual decrease as the dose increased, but remaining at satisfactory levels. Conclusion Ionizing radiation promoted structural changes in the tetanus toxin such as fragmentation and/or aggregation and attenuation of enzymatic activity as the dose increased, but antigenic recognition of the toxin remained at good levels indicating its possible use as an immunogen. However, studies of enzymatic activity of tetanus toxin irradiated with doses above 8 kGy should be further analyzed.Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP/UNESP)2021-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992021000100310Journal of Venomous Animals and Toxins including Tropical Diseases v.27 2021reponame:The Journal of venomous animals and toxins including tropical diseases (Online)instname:Universidade Estadual Paulista (UNESP)instacron:UNESP10.1590/1678-9199-jvatitd-2020-0140info:eu-repo/semantics/openAccessSartori,Giselle PacificoCosta,Andréa daMacarini,Fernanda Lúcio dos SantosMariano,Douglas Oscar CeolinPimenta,Daniel CarvalhoSpencer,Patrick JackNali,Luiz Henrique da SilvaGalisteo Jr.,Andrés Jimenezeng2021-04-27T00:00:00Zoai:scielo:S1678-91992021000100310Revistahttp://www.scielo.br/jvatitdPUBhttps://old.scielo.br/oai/scielo-oai.php||editorial@jvat.org.br1678-91991678-9180opendoar:2021-04-27T00:00The Journal of venomous animals and toxins including tropical diseases (Online) - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation |
title |
Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation |
spellingShingle |
Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation Sartori,Giselle Pacifico Radiation Fragmentation Enzymatic activity Gamma rays |
title_short |
Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation |
title_full |
Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation |
title_fullStr |
Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation |
title_full_unstemmed |
Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation |
title_sort |
Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation |
author |
Sartori,Giselle Pacifico |
author_facet |
Sartori,Giselle Pacifico Costa,Andréa da Macarini,Fernanda Lúcio dos Santos Mariano,Douglas Oscar Ceolin Pimenta,Daniel Carvalho Spencer,Patrick Jack Nali,Luiz Henrique da Silva Galisteo Jr.,Andrés Jimenez |
author_role |
author |
author2 |
Costa,Andréa da Macarini,Fernanda Lúcio dos Santos Mariano,Douglas Oscar Ceolin Pimenta,Daniel Carvalho Spencer,Patrick Jack Nali,Luiz Henrique da Silva Galisteo Jr.,Andrés Jimenez |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Sartori,Giselle Pacifico Costa,Andréa da Macarini,Fernanda Lúcio dos Santos Mariano,Douglas Oscar Ceolin Pimenta,Daniel Carvalho Spencer,Patrick Jack Nali,Luiz Henrique da Silva Galisteo Jr.,Andrés Jimenez |
dc.subject.por.fl_str_mv |
Radiation Fragmentation Enzymatic activity Gamma rays |
topic |
Radiation Fragmentation Enzymatic activity Gamma rays |
description |
Abstract Background Tetanus toxin blocks the release of the inhibitory neurotransmitters in the central nervous system and causes tetanus and its main form of prevention is through vaccination. The vaccine is produced by inactivation of tetanus toxin with formaldehyde, which may cause side effects. An alternative way is the use of ionizing radiation for inactivation of the toxin and also to improve the potential immunogenic response and to reduce the post-vaccination side effects. Therefore, the aim of this study was to characterize the tetanus toxin structure after different doses of ionizing radiation of 60Co. Methods Irradiated and native tetanus toxin was characterized by SDS PAGE in reducing and non-reducing conditions and MALD-TOF. Enzymatic activity was measured by FRET substrate. Also, antigenic properties were assessed by ELISA and Western Blot data. Results Characterization analysis revealed gradual modification on the tetanus toxin structure according to doses increase. Also, fragmentation and possible aggregations of the protein fragments were observed in higher doses. In the analysis of peptide preservation by enzymatic digestion and mass spectrometry, there was a slight modification in the identification up to the dose of 4 kGy. At subsequent doses, peptide identification was minimal. The analysis of the enzymatic activity by fluorescence showed 35 % attenuation in the activity even at higher doses. In the antigenic evaluation, anti-tetanus toxin antibodies were detected against the irradiated toxins at the different doses, with a gradual decrease as the dose increased, but remaining at satisfactory levels. Conclusion Ionizing radiation promoted structural changes in the tetanus toxin such as fragmentation and/or aggregation and attenuation of enzymatic activity as the dose increased, but antigenic recognition of the toxin remained at good levels indicating its possible use as an immunogen. However, studies of enzymatic activity of tetanus toxin irradiated with doses above 8 kGy should be further analyzed. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-01-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992021000100310 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992021000100310 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/1678-9199-jvatitd-2020-0140 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP/UNESP) |
publisher.none.fl_str_mv |
Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP/UNESP) |
dc.source.none.fl_str_mv |
Journal of Venomous Animals and Toxins including Tropical Diseases v.27 2021 reponame:The Journal of venomous animals and toxins including tropical diseases (Online) instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
The Journal of venomous animals and toxins including tropical diseases (Online) |
collection |
The Journal of venomous animals and toxins including tropical diseases (Online) |
repository.name.fl_str_mv |
The Journal of venomous animals and toxins including tropical diseases (Online) - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
||editorial@jvat.org.br |
_version_ |
1748958541047136256 |