Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system

Detalhes bibliográficos
Autor(a) principal: Silva Junior,Haroldo Cid da
Data de Publicação: 2016
Outros Autores: Pestana,Cristiane Pinheiro, Galler,Ricardo, Medeiros,Marco Alberto
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Memórias do Instituto Oswaldo Cruz
Texto Completo: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762016000800535
Resumo: The use of recombinant proteins may represent an alternative model to inactivated vaccines against hepatitis A virus (HAV). The present study aimed to express the VP1 protein of HAV in baculovirus expression vector system (BEVS). The VP1 was expressed intracellularly with molecular mass of 35 kDa. The VP1 was detected both in the soluble fraction and in the insoluble fraction of the lysate. The extracellular expression of VP1 was also attempted, but the protein remained inside the cell. To verify if hydrophobic characteristics would also be present in the HAV structural polyprotein, the expression of P1-2A protein was evaluated. The P1-2A polyprotein remained insoluble in the cellular extract, even in the early infection stages. These results suggest that HAV structural proteins are prone to form insoluble aggregates. The low solubility represents a drawback for production of large amounts of HAV proteins in BEVS.
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spelling Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus systembaculovirusexpressionhepatitis Asolubility and vaccineThe use of recombinant proteins may represent an alternative model to inactivated vaccines against hepatitis A virus (HAV). The present study aimed to express the VP1 protein of HAV in baculovirus expression vector system (BEVS). The VP1 was expressed intracellularly with molecular mass of 35 kDa. The VP1 was detected both in the soluble fraction and in the insoluble fraction of the lysate. The extracellular expression of VP1 was also attempted, but the protein remained inside the cell. To verify if hydrophobic characteristics would also be present in the HAV structural polyprotein, the expression of P1-2A protein was evaluated. The P1-2A polyprotein remained insoluble in the cellular extract, even in the early infection stages. These results suggest that HAV structural proteins are prone to form insoluble aggregates. The low solubility represents a drawback for production of large amounts of HAV proteins in BEVS.Instituto Oswaldo Cruz, Ministério da Saúde2016-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762016000800535Memórias do Instituto Oswaldo Cruz v.111 n.8 2016reponame:Memórias do Instituto Oswaldo Cruzinstname:Fundação Oswaldo Cruzinstacron:FIOCRUZ10.1590/0074-02760160153info:eu-repo/semantics/openAccessSilva Junior,Haroldo Cid daPestana,Cristiane PinheiroGaller,RicardoMedeiros,Marco Albertoeng2020-04-25T17:52:27Zhttp://www.scielo.br/oai/scielo-oai.php0074-02761678-8060opendoar:null2020-04-26 02:21:20.0Memórias do Instituto Oswaldo Cruz - Fundação Oswaldo Cruztrue
dc.title.none.fl_str_mv Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system
title Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system
spellingShingle Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system
Silva Junior,Haroldo Cid da
baculovirus
expression
hepatitis A
solubility and vaccine
title_short Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system
title_full Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system
title_fullStr Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system
title_full_unstemmed Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system
title_sort Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system
author Silva Junior,Haroldo Cid da
author_facet Silva Junior,Haroldo Cid da
Pestana,Cristiane Pinheiro
Galler,Ricardo
Medeiros,Marco Alberto
author_role author
author2 Pestana,Cristiane Pinheiro
Galler,Ricardo
Medeiros,Marco Alberto
author2_role author
author
author
dc.contributor.author.fl_str_mv Silva Junior,Haroldo Cid da
Pestana,Cristiane Pinheiro
Galler,Ricardo
Medeiros,Marco Alberto
dc.subject.por.fl_str_mv baculovirus
expression
hepatitis A
solubility and vaccine
topic baculovirus
expression
hepatitis A
solubility and vaccine
dc.description.none.fl_txt_mv The use of recombinant proteins may represent an alternative model to inactivated vaccines against hepatitis A virus (HAV). The present study aimed to express the VP1 protein of HAV in baculovirus expression vector system (BEVS). The VP1 was expressed intracellularly with molecular mass of 35 kDa. The VP1 was detected both in the soluble fraction and in the insoluble fraction of the lysate. The extracellular expression of VP1 was also attempted, but the protein remained inside the cell. To verify if hydrophobic characteristics would also be present in the HAV structural polyprotein, the expression of P1-2A protein was evaluated. The P1-2A polyprotein remained insoluble in the cellular extract, even in the early infection stages. These results suggest that HAV structural proteins are prone to form insoluble aggregates. The low solubility represents a drawback for production of large amounts of HAV proteins in BEVS.
description The use of recombinant proteins may represent an alternative model to inactivated vaccines against hepatitis A virus (HAV). The present study aimed to express the VP1 protein of HAV in baculovirus expression vector system (BEVS). The VP1 was expressed intracellularly with molecular mass of 35 kDa. The VP1 was detected both in the soluble fraction and in the insoluble fraction of the lysate. The extracellular expression of VP1 was also attempted, but the protein remained inside the cell. To verify if hydrophobic characteristics would also be present in the HAV structural polyprotein, the expression of P1-2A protein was evaluated. The P1-2A polyprotein remained insoluble in the cellular extract, even in the early infection stages. These results suggest that HAV structural proteins are prone to form insoluble aggregates. The low solubility represents a drawback for production of large amounts of HAV proteins in BEVS.
publishDate 2016
dc.date.none.fl_str_mv 2016-08-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762016000800535
url http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762016000800535
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/0074-02760160153
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Instituto Oswaldo Cruz, Ministério da Saúde
publisher.none.fl_str_mv Instituto Oswaldo Cruz, Ministério da Saúde
dc.source.none.fl_str_mv Memórias do Instituto Oswaldo Cruz v.111 n.8 2016
reponame:Memórias do Instituto Oswaldo Cruz
instname:Fundação Oswaldo Cruz
instacron:FIOCRUZ
reponame_str Memórias do Instituto Oswaldo Cruz
collection Memórias do Instituto Oswaldo Cruz
instname_str Fundação Oswaldo Cruz
instacron_str FIOCRUZ
institution FIOCRUZ
repository.name.fl_str_mv Memórias do Instituto Oswaldo Cruz - Fundação Oswaldo Cruz
repository.mail.fl_str_mv
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