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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer International Publishing
2013
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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. |
format | Online Article Text |
id | pubmed-4255388 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Springer International Publishing |
record_format | MEDLINE/PubMed |
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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