The F309S mutation increases factor VIII secretion in human cell line
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Revista brasileira de hematologia e hemoterapia (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-84842016000200135 |
Resumo: | ABSTRACT OBJECTIVES: The capacity of a human cell line to secrete recombinant factor VIII with a F309S point mutation was investigated, as was the effect of the addition of chemical chaperones (betaine and sodium-4-phenylbutyrate) on the secretion of factor VIII. METHODS: This work used a vector with a F309S mutation in the A1 domain to investigate FVIII production in the HEK 293 human cell line. Factor VIII activity was measured by chromogenic assay. Furthermore, the effects of chemical drugs on the culture were evaluated. RESULTS: The addition of the F309S mutation to a previously described FVIII variant increased FVIII secretion by 4.5 fold. Moreover, the addition of betaine or sodium-4-phenylbutyrate increased the secretion rate of FVIIIΔB proteins in HEK 293 cells, but the same effect was not seen for FVIIIΔB-F309S indicating that all the recombinant protein produced had been efficiently secreted. CONCLUSION: Bioengineering factor VIII expressed in human cells may lead to an efficient production of recombinant factor VIII and contribute toward low-cost coagulation factor replacement therapy for hemophilia A. FVIII-F309S produced in human cells can be effective in vivo. |
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Revista brasileira de hematologia e hemoterapia (Online) |
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The F309S mutation increases factor VIII secretion in human cell lineHEK 293 cellsRecombinant factor VIIIF309S mutationBetaineSodium-4-phenylbutyrateABSTRACT OBJECTIVES: The capacity of a human cell line to secrete recombinant factor VIII with a F309S point mutation was investigated, as was the effect of the addition of chemical chaperones (betaine and sodium-4-phenylbutyrate) on the secretion of factor VIII. METHODS: This work used a vector with a F309S mutation in the A1 domain to investigate FVIII production in the HEK 293 human cell line. Factor VIII activity was measured by chromogenic assay. Furthermore, the effects of chemical drugs on the culture were evaluated. RESULTS: The addition of the F309S mutation to a previously described FVIII variant increased FVIII secretion by 4.5 fold. Moreover, the addition of betaine or sodium-4-phenylbutyrate increased the secretion rate of FVIIIΔB proteins in HEK 293 cells, but the same effect was not seen for FVIIIΔB-F309S indicating that all the recombinant protein produced had been efficiently secreted. CONCLUSION: Bioengineering factor VIII expressed in human cells may lead to an efficient production of recombinant factor VIII and contribute toward low-cost coagulation factor replacement therapy for hemophilia A. FVIII-F309S produced in human cells can be effective in vivo.Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular2016-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-84842016000200135Revista Brasileira de Hematologia e Hemoterapia v.38 n.2 2016reponame:Revista brasileira de hematologia e hemoterapia (Online)instname:Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular (ABHHTC)instacron:ABHHTC10.1016/j.bjhh.2016.04.002info:eu-repo/semantics/openAccessFantacini,Daianne Maciely CarvalhoFontes,Aparecida MariaAbreu Neto,Mário Soares deCovas,Dimas TadeuPicanço-Castro,Virgíniaeng2016-06-29T00:00:00Zoai:scielo:S1516-84842016000200135Revistahttp://www.rbhh.org/pt/archivo/https://old.scielo.br/oai/scielo-oai.phpsbhh@terra.com.br||secretaria@rbhh.org1806-08701516-8484opendoar:2016-06-29T00:00Revista brasileira de hematologia e hemoterapia (Online) - Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular (ABHHTC)false |
dc.title.none.fl_str_mv |
The F309S mutation increases factor VIII secretion in human cell line |
title |
The F309S mutation increases factor VIII secretion in human cell line |
spellingShingle |
The F309S mutation increases factor VIII secretion in human cell line Fantacini,Daianne Maciely Carvalho HEK 293 cells Recombinant factor VIII F309S mutation Betaine Sodium-4-phenylbutyrate |
title_short |
The F309S mutation increases factor VIII secretion in human cell line |
title_full |
The F309S mutation increases factor VIII secretion in human cell line |
title_fullStr |
The F309S mutation increases factor VIII secretion in human cell line |
title_full_unstemmed |
The F309S mutation increases factor VIII secretion in human cell line |
title_sort |
The F309S mutation increases factor VIII secretion in human cell line |
author |
Fantacini,Daianne Maciely Carvalho |
author_facet |
Fantacini,Daianne Maciely Carvalho Fontes,Aparecida Maria Abreu Neto,Mário Soares de Covas,Dimas Tadeu Picanço-Castro,Virgínia |
author_role |
author |
author2 |
Fontes,Aparecida Maria Abreu Neto,Mário Soares de Covas,Dimas Tadeu Picanço-Castro,Virgínia |
author2_role |
author author author author |
dc.contributor.author.fl_str_mv |
Fantacini,Daianne Maciely Carvalho Fontes,Aparecida Maria Abreu Neto,Mário Soares de Covas,Dimas Tadeu Picanço-Castro,Virgínia |
dc.subject.por.fl_str_mv |
HEK 293 cells Recombinant factor VIII F309S mutation Betaine Sodium-4-phenylbutyrate |
topic |
HEK 293 cells Recombinant factor VIII F309S mutation Betaine Sodium-4-phenylbutyrate |
description |
ABSTRACT OBJECTIVES: The capacity of a human cell line to secrete recombinant factor VIII with a F309S point mutation was investigated, as was the effect of the addition of chemical chaperones (betaine and sodium-4-phenylbutyrate) on the secretion of factor VIII. METHODS: This work used a vector with a F309S mutation in the A1 domain to investigate FVIII production in the HEK 293 human cell line. Factor VIII activity was measured by chromogenic assay. Furthermore, the effects of chemical drugs on the culture were evaluated. RESULTS: The addition of the F309S mutation to a previously described FVIII variant increased FVIII secretion by 4.5 fold. Moreover, the addition of betaine or sodium-4-phenylbutyrate increased the secretion rate of FVIIIΔB proteins in HEK 293 cells, but the same effect was not seen for FVIIIΔB-F309S indicating that all the recombinant protein produced had been efficiently secreted. CONCLUSION: Bioengineering factor VIII expressed in human cells may lead to an efficient production of recombinant factor VIII and contribute toward low-cost coagulation factor replacement therapy for hemophilia A. FVIII-F309S produced in human cells can be effective in vivo. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-06-01 |
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://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-84842016000200135 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-84842016000200135 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1016/j.bjhh.2016.04.002 |
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 |
Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular |
publisher.none.fl_str_mv |
Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular |
dc.source.none.fl_str_mv |
Revista Brasileira de Hematologia e Hemoterapia v.38 n.2 2016 reponame:Revista brasileira de hematologia e hemoterapia (Online) instname:Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular (ABHHTC) instacron:ABHHTC |
instname_str |
Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular (ABHHTC) |
instacron_str |
ABHHTC |
institution |
ABHHTC |
reponame_str |
Revista brasileira de hematologia e hemoterapia (Online) |
collection |
Revista brasileira de hematologia e hemoterapia (Online) |
repository.name.fl_str_mv |
Revista brasileira de hematologia e hemoterapia (Online) - Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular (ABHHTC) |
repository.mail.fl_str_mv |
sbhh@terra.com.br||secretaria@rbhh.org |
_version_ |
1754213112812142592 |