Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60

Detalhes bibliográficos
Autor(a) principal: SARTORI, GISELLE P.
Data de Publicação: 2020
Outros Autores: COSTA, ANDREA, MACARINI, FERNANDA L.S., MARIANO, DOUGLAS O.C., PIMENTA, DANIEL C., SPENCER, PATRICK J., GALISTEO JUNIOR, ANDRES J., INTERNATIONAL NUCLEAR ATLANTIC CONFERENCE
Tipo de documento: Artigo de conferência
Título da fonte: Repositório Institucional do IPEN
Texto Completo: http://repositorio.ipen.br/handle/123456789/30616
Resumo: Tetanus is a neurological disease which blocks the inhibitory neurotransmitters liberation. Also treatment does not cure the disease, and its main form of prevention is through vaccination. The vaccine is produced by the inactivation of tetanic toxin (TeNT) 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 pos-vaccination side effects. Therefore, the aim of this study was to characterize the TeNT structure after different doses of ionizing radiation of Cobalt 60, and also to assess the enzymatic activity after the radiation. SDS PAGE and MALDI-TOF analysis revealed gradual modification on the TeNT 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 recognition up to the dose of 4 kGy at subsequent doses, recognition was minimal. The analysis of the enzymatic activity by fluorescence showed a 35 % attenuation in the activity even at higher doses. In the antigenic evaluation performed by ELISA and Western Blot, anti-TeNT antibodies were detected against the irradiated toxins at the different doses, with a gradual decrease as the dose increased, but remaining at satisfactory levels, indicating the possibility of possible use as an immunogen, however, studies of enzymatic activity on higher should be further analyzed.
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spelling 2020-01-10T12:14:39Z2020-01-10T12:14:39ZOctober 21-25, 2019http://repositorio.ipen.br/handle/123456789/306160000-0001-8949-7735Tetanus is a neurological disease which blocks the inhibitory neurotransmitters liberation. Also treatment does not cure the disease, and its main form of prevention is through vaccination. The vaccine is produced by the inactivation of tetanic toxin (TeNT) 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 pos-vaccination side effects. Therefore, the aim of this study was to characterize the TeNT structure after different doses of ionizing radiation of Cobalt 60, and also to assess the enzymatic activity after the radiation. SDS PAGE and MALDI-TOF analysis revealed gradual modification on the TeNT 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 recognition up to the dose of 4 kGy at subsequent doses, recognition was minimal. The analysis of the enzymatic activity by fluorescence showed a 35 % attenuation in the activity even at higher doses. In the antigenic evaluation performed by ELISA and Western Blot, anti-TeNT antibodies were detected against the irradiated toxins at the different doses, with a gradual decrease as the dose increased, but remaining at satisfactory levels, indicating the possibility of possible use as an immunogen, however, studies of enzymatic activity on higher should be further analyzed.Submitted by Celia Satomi Uehara (celia.u-topservice@ipen.br) on 2020-01-10T12:14:39Z No. of bitstreams: 1 26265.pdf: 700443 bytes, checksum: 221cb862221da3f775501b8861790e79 (MD5)Made available in DSpace on 2020-01-10T12:14:39Z (GMT). No. of bitstreams: 1 26265.pdf: 700443 bytes, checksum: 221cb862221da3f775501b8861790e79 (MD5)2191-2203Associa????o Brasileira de Energia Nuclearcobalt 60detoxificationenzyme activitygamma radiationirradiationneurologyradiation dose unitstetanustoxinsvaccinesCharacterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectINACIRio de JaneiroSantos, SP910600SARTORI, GISELLE P.COSTA, ANDREAMACARINI, FERNANDA L.S.MARIANO, DOUGLAS O.C.PIMENTA, DANIEL C.SPENCER, PATRICK J.GALISTEO JUNIOR, ANDRES J.INTERNATIONAL NUCLEAR ATLANTIC CONFERENCEinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional do IPENinstname:Instituto de Pesquisas Energéticas e Nucleares (IPEN)instacron:IPEN262652019SPENCER, PATRICK J.20-01Proceedings910SPENCER, PATRICK J.:910:820:NORIGINAL26265.pdf26265.pdfapplication/pdf700443http://repositorio.ipen.br/bitstream/123456789/30616/1/26265.pdf221cb862221da3f775501b8861790e79MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ipen.br/bitstream/123456789/30616/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52123456789/306162020-03-23 18:48:38.172oai:repositorio.ipen.br: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Repositório InstitucionalPUBhttp://repositorio.ipen.br/oai/requestbibl@ipen.bropendoar:45102020-03-23T18:48:38Repositório Institucional do IPEN - Instituto de Pesquisas Energéticas e Nucleares (IPEN)false
dc.title.pt_BR.fl_str_mv Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60
title Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60
spellingShingle Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60
SARTORI, GISELLE P.
cobalt 60
detoxification
enzyme activity
gamma radiation
irradiation
neurology
radiation dose units
tetanus
toxins
vaccines
title_short Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60
title_full Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60
title_fullStr Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60
title_full_unstemmed Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60
title_sort Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60
author SARTORI, GISELLE P.
author_facet SARTORI, GISELLE P.
COSTA, ANDREA
MACARINI, FERNANDA L.S.
MARIANO, DOUGLAS O.C.
PIMENTA, DANIEL C.
SPENCER, PATRICK J.
GALISTEO JUNIOR, ANDRES J.
INTERNATIONAL NUCLEAR ATLANTIC CONFERENCE
author_role author
author2 COSTA, ANDREA
MACARINI, FERNANDA L.S.
MARIANO, DOUGLAS O.C.
PIMENTA, DANIEL C.
SPENCER, PATRICK J.
GALISTEO JUNIOR, ANDRES J.
INTERNATIONAL NUCLEAR ATLANTIC CONFERENCE
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv SARTORI, GISELLE P.
COSTA, ANDREA
MACARINI, FERNANDA L.S.
MARIANO, DOUGLAS O.C.
PIMENTA, DANIEL C.
SPENCER, PATRICK J.
GALISTEO JUNIOR, ANDRES J.
INTERNATIONAL NUCLEAR ATLANTIC CONFERENCE
dc.subject.por.fl_str_mv cobalt 60
detoxification
enzyme activity
gamma radiation
irradiation
neurology
radiation dose units
tetanus
toxins
vaccines
topic cobalt 60
detoxification
enzyme activity
gamma radiation
irradiation
neurology
radiation dose units
tetanus
toxins
vaccines
description Tetanus is a neurological disease which blocks the inhibitory neurotransmitters liberation. Also treatment does not cure the disease, and its main form of prevention is through vaccination. The vaccine is produced by the inactivation of tetanic toxin (TeNT) 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 pos-vaccination side effects. Therefore, the aim of this study was to characterize the TeNT structure after different doses of ionizing radiation of Cobalt 60, and also to assess the enzymatic activity after the radiation. SDS PAGE and MALDI-TOF analysis revealed gradual modification on the TeNT 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 recognition up to the dose of 4 kGy at subsequent doses, recognition was minimal. The analysis of the enzymatic activity by fluorescence showed a 35 % attenuation in the activity even at higher doses. In the antigenic evaluation performed by ELISA and Western Blot, anti-TeNT antibodies were detected against the irradiated toxins at the different doses, with a gradual decrease as the dose increased, but remaining at satisfactory levels, indicating the possibility of possible use as an immunogen, however, studies of enzymatic activity on higher should be further analyzed.
publishDate 2020
dc.date.evento.pt_BR.fl_str_mv October 21-25, 2019
dc.date.accessioned.fl_str_mv 2020-01-10T12:14:39Z
dc.date.available.fl_str_mv 2020-01-10T12:14:39Z
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identifier_str_mv 0000-0001-8949-7735
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publisher.none.fl_str_mv Associa????o Brasileira de Energia Nuclear
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