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Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line

Hek-293 cell line presents good production platform for recombinant therapeutic proteins, however little is known about the components that contribute to the cellular control of recombinant protein production. In this study, we generated a Hek-293 producing recombinant factor VIII (FVIII) and we eva...

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Autores principales: Rodrigues, Evandra Strazza, Picanço-Castro, Virgínia, Espanhol, Marta Regina, de Andrade, Luiz Alberto Martins, Palma, Patricia Vianna Bonini, Kashima, Simone, Fontes, Aparecida Maria, Covas, Dimas Tadeu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4255388/
https://www.ncbi.nlm.nih.gov/pubmed/25530931
http://dx.doi.org/10.1186/2193-1801-2-328
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author Rodrigues, Evandra Strazza
Picanço-Castro, Virgínia
Espanhol, Marta Regina
de Andrade, Luiz Alberto Martins
Palma, Patricia Vianna Bonini
Kashima, Simone
Fontes, Aparecida Maria
Covas, Dimas Tadeu
author_facet Rodrigues, Evandra Strazza
Picanço-Castro, Virgínia
Espanhol, Marta Regina
de Andrade, Luiz Alberto Martins
Palma, Patricia Vianna Bonini
Kashima, Simone
Fontes, Aparecida Maria
Covas, Dimas Tadeu
author_sort Rodrigues, Evandra Strazza
collection PubMed
description Hek-293 cell line presents good production platform for recombinant therapeutic proteins, however little is known about the components that contribute to the cellular control of recombinant protein production. In this study, we generated a Hek-293 producing recombinant factor VIII (FVIII) and we evaluated the immunoglobulin-binding protein (BiP) and phytanoil-CoA α-hydroxylase (PAHX) expression levels which are known for diminishing FVIII production. Our analyses showed that the recombinant cell population expresses 3.1 ± 1.4 fold of BIP mRNA (P = 0.0054) and 97.8 ± 0.5 fold of PAHX mRNA (P = 0.0016) compared to nontransduced cells. The amount of these proteins was inversely correlated to the secreted FVIII. In conclusion, BIP and PAHX expression are augmented in human cells producing FVIII and they antagonize the amount of therapeutic factor VIII in the cell culture. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/2193-1801-2-328) contains supplementary material, which is available to authorized users.
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spelling pubmed-42553882014-12-19 Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line Rodrigues, Evandra Strazza Picanço-Castro, Virgínia Espanhol, Marta Regina de Andrade, Luiz Alberto Martins Palma, Patricia Vianna Bonini Kashima, Simone Fontes, Aparecida Maria Covas, Dimas Tadeu Springerplus Research Hek-293 cell line presents good production platform for recombinant therapeutic proteins, however little is known about the components that contribute to the cellular control of recombinant protein production. In this study, we generated a Hek-293 producing recombinant factor VIII (FVIII) and we evaluated the immunoglobulin-binding protein (BiP) and phytanoil-CoA α-hydroxylase (PAHX) expression levels which are known for diminishing FVIII production. Our analyses showed that the recombinant cell population expresses 3.1 ± 1.4 fold of BIP mRNA (P = 0.0054) and 97.8 ± 0.5 fold of PAHX mRNA (P = 0.0016) compared to nontransduced cells. The amount of these proteins was inversely correlated to the secreted FVIII. In conclusion, BIP and PAHX expression are augmented in human cells producing FVIII and they antagonize the amount of therapeutic factor VIII in the cell culture. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/2193-1801-2-328) contains supplementary material, which is available to authorized users. Springer International Publishing 2013-07-18 /pmc/articles/PMC4255388/ /pubmed/25530931 http://dx.doi.org/10.1186/2193-1801-2-328 Text en © Rodrigues et al.; licensee Springer. 2013 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Rodrigues, Evandra Strazza
Picanço-Castro, Virgínia
Espanhol, Marta Regina
de Andrade, Luiz Alberto Martins
Palma, Patricia Vianna Bonini
Kashima, Simone
Fontes, Aparecida Maria
Covas, Dimas Tadeu
Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line
title Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line
title_full Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line
title_fullStr Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line
title_full_unstemmed Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line
title_short Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line
title_sort quantitative correlation between transcriptional levels of er chaperone, peroximal protein and fviii productivity in human hek-293 cell line
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4255388/
https://www.ncbi.nlm.nih.gov/pubmed/25530931
http://dx.doi.org/10.1186/2193-1801-2-328
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