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Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β

A CHO cell clone (CHO-PoIFN-β) with stable porcine IFN-β expression under control of CMV promoter was selected under G418 pressure. In a 25 cm(2) cell culture flask (5 ml culture medium), the cumulative protein yield of recombinant PoIFN-β reached 2.3 × 10(6) IU/ml. This cells clone maintained stabl...

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Autores principales: Chen, Weiye, Cao, Wenyan, Zhao, Huijun, Hu, Qianqian, Qu, Linmao, Hu, Sen, Ge, Jinying, Wen, Zhiyuan, Wang, Xijun, Li, HaoBo, Huang, Kehe, Bu, Zhigao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Published by Elsevier Ltd. 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7128424/
https://www.ncbi.nlm.nih.gov/pubmed/21459017
http://dx.doi.org/10.1016/j.cyto.2010.12.002
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author Chen, Weiye
Cao, Wenyan
Zhao, Huijun
Hu, Qianqian
Qu, Linmao
Hu, Sen
Ge, Jinying
Wen, Zhiyuan
Wang, Xijun
Li, HaoBo
Huang, Kehe
Bu, Zhigao
author_facet Chen, Weiye
Cao, Wenyan
Zhao, Huijun
Hu, Qianqian
Qu, Linmao
Hu, Sen
Ge, Jinying
Wen, Zhiyuan
Wang, Xijun
Li, HaoBo
Huang, Kehe
Bu, Zhigao
author_sort Chen, Weiye
collection PubMed
description A CHO cell clone (CHO-PoIFN-β) with stable porcine IFN-β expression under control of CMV promoter was selected under G418 pressure. In a 25 cm(2) cell culture flask (5 ml culture medium), the cumulative protein yield of recombinant PoIFN-β reached 2.3 × 10(6) IU/ml. This cells clone maintained stable expression for at least 20 generations even in the absence of G418 selection pressure. The expressed recombinant PoIFN-β could induce the expression of porcine Mx protein in PK15 cells, and activate the chicken Mx promoter-controlled luciferase reporter gene expression, confirming that the recombinant PoIFN-β has the biological activity of natural porcine type-I interferon. In addition, the recombinant PoIFN-β fully protected PK15 cells against 1000 TCID(50) of porcine transmissible gastroenteritis virus and pseudo-rabies virus infection, demonstrating its high potential in therapeutic applications. This is the first report of establishing a mammalian cell line with stable expression of porcine IFN-β.
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spelling pubmed-71284242020-04-08 Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β Chen, Weiye Cao, Wenyan Zhao, Huijun Hu, Qianqian Qu, Linmao Hu, Sen Ge, Jinying Wen, Zhiyuan Wang, Xijun Li, HaoBo Huang, Kehe Bu, Zhigao Cytokine Article A CHO cell clone (CHO-PoIFN-β) with stable porcine IFN-β expression under control of CMV promoter was selected under G418 pressure. In a 25 cm(2) cell culture flask (5 ml culture medium), the cumulative protein yield of recombinant PoIFN-β reached 2.3 × 10(6) IU/ml. This cells clone maintained stable expression for at least 20 generations even in the absence of G418 selection pressure. The expressed recombinant PoIFN-β could induce the expression of porcine Mx protein in PK15 cells, and activate the chicken Mx promoter-controlled luciferase reporter gene expression, confirming that the recombinant PoIFN-β has the biological activity of natural porcine type-I interferon. In addition, the recombinant PoIFN-β fully protected PK15 cells against 1000 TCID(50) of porcine transmissible gastroenteritis virus and pseudo-rabies virus infection, demonstrating its high potential in therapeutic applications. This is the first report of establishing a mammalian cell line with stable expression of porcine IFN-β. Published by Elsevier Ltd. 2011-06 2011-04-01 /pmc/articles/PMC7128424/ /pubmed/21459017 http://dx.doi.org/10.1016/j.cyto.2010.12.002 Text en Copyright © 2010 Published by Elsevier Ltd. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Chen, Weiye
Cao, Wenyan
Zhao, Huijun
Hu, Qianqian
Qu, Linmao
Hu, Sen
Ge, Jinying
Wen, Zhiyuan
Wang, Xijun
Li, HaoBo
Huang, Kehe
Bu, Zhigao
Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β
title Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β
title_full Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β
title_fullStr Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β
title_full_unstemmed Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β
title_short Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β
title_sort establishment of a stable cho cell line with high level expression of recombinant porcine ifn-β
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7128424/
https://www.ncbi.nlm.nih.gov/pubmed/21459017
http://dx.doi.org/10.1016/j.cyto.2010.12.002
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