Conventional and volatile salts as precipitating agents for recombinant antiviral protein
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Bioscience journal (Online) |
Texto Completo: | https://seer.ufu.br/index.php/biosciencejournal/article/view/36729 |
Resumo: | Recombinant proteins expressed in cell culture have been shown to be relevant in the biopharmaceutical production focusing human health. The current work investigated the precipitation process of recAVLOEc protein, synthesized by E. coli BL21 (DE3) pLysS cells. The system is used for the AVLO expression that shown antiviral activity and it was found in the hemolymph of Lonomia obliqua caterpillar. The precipitation was conducted by the use of conventional salts (ammonium sulfate and sodium sulfate) and the volatile ammonium carbamate salt. Initially, the precipitated protein obtained from bacterial lysate was added to L929 cells to evaluate the cytotoxic effect; and besides Vero cells were infected with measles virus to verify the antiviral action of the precipitated recombinant protein. Toxic effect on the culture of L929 cells was observed for the precipitate obtained by the use of ammonium sulfate and sodium sulfate. In addition, tests in L929 cell cultures infected with EMC virus showed that samples of precipitated protein by salts did not show antiviral action. In Vero cell cultures, the precipitated protein by sodium sulfate showed antiviral action for measles virus. |
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Conventional and volatile salts as precipitating agents for recombinant antiviral protein Sais convencionais e voláteis como agentes de precipitação para proteína recombinante com ação antiviralMeaslesrecAVLOEcSalting outVolatile saltAgricultural Sciences Recombinant proteins expressed in cell culture have been shown to be relevant in the biopharmaceutical production focusing human health. The current work investigated the precipitation process of recAVLOEc protein, synthesized by E. coli BL21 (DE3) pLysS cells. The system is used for the AVLO expression that shown antiviral activity and it was found in the hemolymph of Lonomia obliqua caterpillar. The precipitation was conducted by the use of conventional salts (ammonium sulfate and sodium sulfate) and the volatile ammonium carbamate salt. Initially, the precipitated protein obtained from bacterial lysate was added to L929 cells to evaluate the cytotoxic effect; and besides Vero cells were infected with measles virus to verify the antiviral action of the precipitated recombinant protein. Toxic effect on the culture of L929 cells was observed for the precipitate obtained by the use of ammonium sulfate and sodium sulfate. In addition, tests in L929 cell cultures infected with EMC virus showed that samples of precipitated protein by salts did not show antiviral action. In Vero cell cultures, the precipitated protein by sodium sulfate showed antiviral action for measles virus.Proteínas recombinantes expressas em culturas celulares têm se mostrado importantes na produção de fármacos de interesse para a saúde humana. Este estudo investigou a precipitação da proteína recAVLOEc, sintetizada por células de E. coli BL21 (DE3) pLysS, utilizadas como sistema de expressão da AVLO, proteína com atividade antiviral, originalmente encontrada na hemolinfa da lagarta Lonomia obliqua. A precipitação foi conduzida por meio do uso de sais convencionais (sulfato de amônio e de sódio) e do sal volátil carbamato de amônio. Inicialmente o precipitado proteico obtido do lisado bacteriano foi administrado em culturas de células L929 para avaliar o efeito citotóxico e posteriormente em células Vero infectadas com o vírus do sarampo, para a verificação da ação antiviral. Um efeito tóxico em culturas de L929 foi observado para os precipitados obtidos pelo uso de sulfato de amônio e de sódio. Testes em culturas de L929 infectadas com o vírus EMC foram também efetuados e as amostras de proteínas precipitadas com os sais convencionais e o sal volátil não resultaram em ação antiviral. Em culturas de células Vero, o uso do sulfato e sódio como agente de precipitação das proteínas contidas no lisado bacteriano resultou em ação antiviral para o sarampo.EDUFU2017-07-25info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://seer.ufu.br/index.php/biosciencejournal/article/view/3672910.14393/BJ-v33n4a2017-36729Bioscience Journal ; Vol. 33 No. 4 (2017): July/Aug.; 1048-1053Bioscience Journal ; v. 33 n. 4 (2017): July/Aug.; 1048-10531981-3163reponame:Bioscience journal (Online)instname:Universidade Federal de Uberlândia (UFU)instacron:UFUenghttps://seer.ufu.br/index.php/biosciencejournal/article/view/36729/20674Brazil; ContemporaryCopyright (c) 2017 Kamilla Alves Carvalho, Dalton Giovanni Nogueira da Silva, Ronaldo Zucatelli Mendonça, Erika Ohta Watanabe, Fabiana Regina Xavier Batistahttps://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessCarvalho, Kamilla AlvesNogueira da Silva, Dalton GiovanniMendonça, Ronaldo ZucatelliWatanabe, Erika OhtaXavier Batista, Fabiana Regina2022-02-14T02:19:41Zoai:ojs.www.seer.ufu.br:article/36729Revistahttps://seer.ufu.br/index.php/biosciencejournalPUBhttps://seer.ufu.br/index.php/biosciencejournal/oaibiosciencej@ufu.br||1981-31631516-3725opendoar:2022-02-14T02:19:41Bioscience journal (Online) - Universidade Federal de Uberlândia (UFU)false |
dc.title.none.fl_str_mv |
Conventional and volatile salts as precipitating agents for recombinant antiviral protein Sais convencionais e voláteis como agentes de precipitação para proteína recombinante com ação antiviral |
title |
Conventional and volatile salts as precipitating agents for recombinant antiviral protein |
spellingShingle |
Conventional and volatile salts as precipitating agents for recombinant antiviral protein Carvalho, Kamilla Alves Measles recAVLOEc Salting out Volatile salt Agricultural Sciences |
title_short |
Conventional and volatile salts as precipitating agents for recombinant antiviral protein |
title_full |
Conventional and volatile salts as precipitating agents for recombinant antiviral protein |
title_fullStr |
Conventional and volatile salts as precipitating agents for recombinant antiviral protein |
title_full_unstemmed |
Conventional and volatile salts as precipitating agents for recombinant antiviral protein |
title_sort |
Conventional and volatile salts as precipitating agents for recombinant antiviral protein |
author |
Carvalho, Kamilla Alves |
author_facet |
Carvalho, Kamilla Alves Nogueira da Silva, Dalton Giovanni Mendonça, Ronaldo Zucatelli Watanabe, Erika Ohta Xavier Batista, Fabiana Regina |
author_role |
author |
author2 |
Nogueira da Silva, Dalton Giovanni Mendonça, Ronaldo Zucatelli Watanabe, Erika Ohta Xavier Batista, Fabiana Regina |
author2_role |
author author author author |
dc.contributor.author.fl_str_mv |
Carvalho, Kamilla Alves Nogueira da Silva, Dalton Giovanni Mendonça, Ronaldo Zucatelli Watanabe, Erika Ohta Xavier Batista, Fabiana Regina |
dc.subject.por.fl_str_mv |
Measles recAVLOEc Salting out Volatile salt Agricultural Sciences |
topic |
Measles recAVLOEc Salting out Volatile salt Agricultural Sciences |
description |
Recombinant proteins expressed in cell culture have been shown to be relevant in the biopharmaceutical production focusing human health. The current work investigated the precipitation process of recAVLOEc protein, synthesized by E. coli BL21 (DE3) pLysS cells. The system is used for the AVLO expression that shown antiviral activity and it was found in the hemolymph of Lonomia obliqua caterpillar. The precipitation was conducted by the use of conventional salts (ammonium sulfate and sodium sulfate) and the volatile ammonium carbamate salt. Initially, the precipitated protein obtained from bacterial lysate was added to L929 cells to evaluate the cytotoxic effect; and besides Vero cells were infected with measles virus to verify the antiviral action of the precipitated recombinant protein. Toxic effect on the culture of L929 cells was observed for the precipitate obtained by the use of ammonium sulfate and sodium sulfate. In addition, tests in L929 cell cultures infected with EMC virus showed that samples of precipitated protein by salts did not show antiviral action. In Vero cell cultures, the precipitated protein by sodium sulfate showed antiviral action for measles virus. |
publishDate |
2017 |
dc.date.none.fl_str_mv |
2017-07-25 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://seer.ufu.br/index.php/biosciencejournal/article/view/36729 10.14393/BJ-v33n4a2017-36729 |
url |
https://seer.ufu.br/index.php/biosciencejournal/article/view/36729 |
identifier_str_mv |
10.14393/BJ-v33n4a2017-36729 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://seer.ufu.br/index.php/biosciencejournal/article/view/36729/20674 |
dc.rights.driver.fl_str_mv |
https://creativecommons.org/licenses/by/4.0 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.coverage.none.fl_str_mv |
Brazil; Contemporary |
dc.publisher.none.fl_str_mv |
EDUFU |
publisher.none.fl_str_mv |
EDUFU |
dc.source.none.fl_str_mv |
Bioscience Journal ; Vol. 33 No. 4 (2017): July/Aug.; 1048-1053 Bioscience Journal ; v. 33 n. 4 (2017): July/Aug.; 1048-1053 1981-3163 reponame:Bioscience journal (Online) instname:Universidade Federal de Uberlândia (UFU) instacron:UFU |
instname_str |
Universidade Federal de Uberlândia (UFU) |
instacron_str |
UFU |
institution |
UFU |
reponame_str |
Bioscience journal (Online) |
collection |
Bioscience journal (Online) |
repository.name.fl_str_mv |
Bioscience journal (Online) - Universidade Federal de Uberlândia (UFU) |
repository.mail.fl_str_mv |
biosciencej@ufu.br|| |
_version_ |
1797069077600010240 |