Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to gamma radiation by Cobalt 60
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , |
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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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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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/conferenceObject |
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conferenceObject |
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http://repositorio.ipen.br/handle/123456789/30616 |
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