Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFRN |
Texto Completo: | https://repositorio.ufrn.br/handle/123456789/32482 |
Resumo: | Visceral leishmaniasis, a disease caused by Leishmania infantum chagasi, represents a major public health problem in many areas of the world. However, there is currently no vaccine for human use. The aim of this work was to purify the 503 antigen of Leishmania i. chagasi directly from unclarified Escherichia coli feedstock through expanded bed adsorption (EBA) chromatography. Batch experiments were performed to optimize the adsorption and elution conditions of the antigen onto a STREAMLINETM Chelating resin using two central composite rotatable designs (CCRD). The results showed that the optimal binding con- ditions of the 503 antigen were pH 8.0 in the presence of 2.4 M NaCl. For the elution of the target protein, the optimized conditions included the presence of 600.0 mM imidazole. The adsorption isothermal data of the 503 antigen were fitted to the Langmuir adsorption isotherm. The EBA experiment successfully recovered 59.2% of the 503 antigen from the unclarified E. coli homogenate with a purification factor of 6.0 |
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Santos, Everaldo Silvino dosSousa Junior, Francisco Caninde deVaz, Michelle Rossana FerreiraPadilha, Carlos Eduardo de AraújoChibério, Abimaelle SilvaMartins, Daniella Regina ArantesMacedo, Gorete Ribeiro de2021-05-10T19:50:34Z2021-05-10T19:50:34Z2015-04-01SOUSA JUNIOR, F. C.; VAZ, M. R. F.; PADILHA, C. E.; CHIBERIO, A. S.; MARTINS, D. R. A.; MACEDO, G. R.; SANTOS, E. S.. Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography. Journal of Chromatography. B (Print), p. 1, 2015. Disponivel em https://www.sciencedirect.com/science/article/abs/pii/S1570023215000719?via%3Dihub. Acesso em: 01 abr. 2021. https://doi.org/10.1016/j.jchromb.2015.01.0311570-0232https://repositorio.ufrn.br/handle/123456789/3248210.1016/j.jchromb.2015.01.031ElsevierAttribution 3.0 Brazilhttp://creativecommons.org/licenses/by/3.0/br/info:eu-repo/semantics/openAccessExpanded bed adsorptionLeishmania infantum chagasiRecombinant protein purificationUnclarified bacterial homogenateVisceral leishmaniasisRecovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatographyinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleVisceral leishmaniasis, a disease caused by Leishmania infantum chagasi, represents a major public health problem in many areas of the world. However, there is currently no vaccine for human use. The aim of this work was to purify the 503 antigen of Leishmania i. chagasi directly from unclarified Escherichia coli feedstock through expanded bed adsorption (EBA) chromatography. Batch experiments were performed to optimize the adsorption and elution conditions of the antigen onto a STREAMLINETM Chelating resin using two central composite rotatable designs (CCRD). The results showed that the optimal binding con- ditions of the 503 antigen were pH 8.0 in the presence of 2.4 M NaCl. For the elution of the target protein, the optimized conditions included the presence of 600.0 mM imidazole. The adsorption isothermal data of the 503 antigen were fitted to the Langmuir adsorption isotherm. The EBA experiment successfully recovered 59.2% of the 503 antigen from the unclarified E. coli homogenate with a purification factor of 6.0engreponame:Repositório Institucional da UFRNinstname:Universidade Federal do Rio Grande do Norte (UFRN)instacron:UFRNORIGINALRecoveryPurificationRecombinant_Santos_2015.pdfRecoveryPurificationRecombinant_Santos_2015.pdfapplication/pdf913543https://repositorio.ufrn.br/bitstream/123456789/32482/1/RecoveryPurificationRecombinant_Santos_2015.pdf08388706a5057e2d75befa37e0f4f4b6MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8914https://repositorio.ufrn.br/bitstream/123456789/32482/2/license_rdf4d2950bda3d176f570a9f8b328dfbbefMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81484https://repositorio.ufrn.br/bitstream/123456789/32482/3/license.txte9597aa2854d128fd968be5edc8a28d9MD53123456789/324822021-05-10 16:50:35.794oai:https://repositorio.ufrn.br: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Repositório de PublicaçõesPUBhttp://repositorio.ufrn.br/oai/opendoar:2021-05-10T19:50:35Repositório Institucional da UFRN - Universidade Federal do Rio Grande do Norte (UFRN)false |
dc.title.pt_BR.fl_str_mv |
Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography |
title |
Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography |
spellingShingle |
Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography Santos, Everaldo Silvino dos Expanded bed adsorption Leishmania infantum chagasi Recombinant protein purification Unclarified bacterial homogenate Visceral leishmaniasis |
title_short |
Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography |
title_full |
Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography |
title_fullStr |
Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography |
title_full_unstemmed |
Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography |
title_sort |
Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography |
author |
Santos, Everaldo Silvino dos |
author_facet |
Santos, Everaldo Silvino dos Sousa Junior, Francisco Caninde de Vaz, Michelle Rossana Ferreira Padilha, Carlos Eduardo de Araújo Chibério, Abimaelle Silva Martins, Daniella Regina Arantes Macedo, Gorete Ribeiro de |
author_role |
author |
author2 |
Sousa Junior, Francisco Caninde de Vaz, Michelle Rossana Ferreira Padilha, Carlos Eduardo de Araújo Chibério, Abimaelle Silva Martins, Daniella Regina Arantes Macedo, Gorete Ribeiro de |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Santos, Everaldo Silvino dos Sousa Junior, Francisco Caninde de Vaz, Michelle Rossana Ferreira Padilha, Carlos Eduardo de Araújo Chibério, Abimaelle Silva Martins, Daniella Regina Arantes Macedo, Gorete Ribeiro de |
dc.subject.por.fl_str_mv |
Expanded bed adsorption Leishmania infantum chagasi Recombinant protein purification Unclarified bacterial homogenate Visceral leishmaniasis |
topic |
Expanded bed adsorption Leishmania infantum chagasi Recombinant protein purification Unclarified bacterial homogenate Visceral leishmaniasis |
description |
Visceral leishmaniasis, a disease caused by Leishmania infantum chagasi, represents a major public health problem in many areas of the world. However, there is currently no vaccine for human use. The aim of this work was to purify the 503 antigen of Leishmania i. chagasi directly from unclarified Escherichia coli feedstock through expanded bed adsorption (EBA) chromatography. Batch experiments were performed to optimize the adsorption and elution conditions of the antigen onto a STREAMLINETM Chelating resin using two central composite rotatable designs (CCRD). The results showed that the optimal binding con- ditions of the 503 antigen were pH 8.0 in the presence of 2.4 M NaCl. For the elution of the target protein, the optimized conditions included the presence of 600.0 mM imidazole. The adsorption isothermal data of the 503 antigen were fitted to the Langmuir adsorption isotherm. The EBA experiment successfully recovered 59.2% of the 503 antigen from the unclarified E. coli homogenate with a purification factor of 6.0 |
publishDate |
2015 |
dc.date.issued.fl_str_mv |
2015-04-01 |
dc.date.accessioned.fl_str_mv |
2021-05-10T19:50:34Z |
dc.date.available.fl_str_mv |
2021-05-10T19:50:34Z |
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 |
SOUSA JUNIOR, F. C.; VAZ, M. R. F.; PADILHA, C. E.; CHIBERIO, A. S.; MARTINS, D. R. A.; MACEDO, G. R.; SANTOS, E. S.. Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography. Journal of Chromatography. B (Print), p. 1, 2015. Disponivel em https://www.sciencedirect.com/science/article/abs/pii/S1570023215000719?via%3Dihub. Acesso em: 01 abr. 2021. https://doi.org/10.1016/j.jchromb.2015.01.031 |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufrn.br/handle/123456789/32482 |
dc.identifier.issn.none.fl_str_mv |
1570-0232 |
dc.identifier.doi.none.fl_str_mv |
10.1016/j.jchromb.2015.01.031 |
identifier_str_mv |
SOUSA JUNIOR, F. C.; VAZ, M. R. F.; PADILHA, C. E.; CHIBERIO, A. S.; MARTINS, D. R. A.; MACEDO, G. R.; SANTOS, E. S.. Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography. Journal of Chromatography. B (Print), p. 1, 2015. Disponivel em https://www.sciencedirect.com/science/article/abs/pii/S1570023215000719?via%3Dihub. Acesso em: 01 abr. 2021. https://doi.org/10.1016/j.jchromb.2015.01.031 1570-0232 10.1016/j.jchromb.2015.01.031 |
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https://repositorio.ufrn.br/handle/123456789/32482 |
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eng |
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eng |
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Attribution 3.0 Brazil http://creativecommons.org/licenses/by/3.0/br/ info:eu-repo/semantics/openAccess |
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Attribution 3.0 Brazil http://creativecommons.org/licenses/by/3.0/br/ |
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openAccess |
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Elsevier |
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Elsevier |
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