Conventional and volatile salts as precipitating agents for recombinant antiviral protein

Detalhes bibliográficos
Autor(a) principal: Carvalho, Kamilla Alves
Data de Publicação: 2017
Outros Autores: Nogueira da Silva, Dalton Giovanni, Mendonça, Ronaldo Zucatelli, Watanabe, Erika Ohta, Xavier Batista, Fabiana Regina
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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spelling 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||
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