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IFN-λ1 in CHO cells: its expression and biological activity

BACKGROUND: Many studies have investigated the characteristics and biological activities of type III interferon (IFN), finding that it has similar features to type I IFN but also unique actions because it is recognized by a different receptor. RESULTS: A full-length recombinant human IFN-λ1 (rhIFN-λ...

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Autores principales: Yuan, Wu-mei, Zhang, Wan-ju, Ma, Fen-lian, Li, Jin-song, Zhang, Qian, Zheng, Li-shu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5709978/
https://www.ncbi.nlm.nih.gov/pubmed/29213292
http://dx.doi.org/10.1186/s11658-017-0057-x
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author Yuan, Wu-mei
Zhang, Wan-ju
Ma, Fen-lian
Li, Jin-song
Zhang, Qian
Zheng, Li-shu
author_facet Yuan, Wu-mei
Zhang, Wan-ju
Ma, Fen-lian
Li, Jin-song
Zhang, Qian
Zheng, Li-shu
author_sort Yuan, Wu-mei
collection PubMed
description BACKGROUND: Many studies have investigated the characteristics and biological activities of type III interferon (IFN), finding that it has similar features to type I IFN but also unique actions because it is recognized by a different receptor. RESULTS: A full-length recombinant human IFN-λ1 (rhIFN-λ1) cDNA was cloned into the pDF expression vector and stably expressed in Flp-In-CHO cells. After four purification steps (ammonium sulfate precipitation, SP Sepharose chromatography, Blue Sepharose 6 fast flow affinity chromatography and molecular sieve chromatography), the rhIFN-λ1 had a purity of about 90% and was found to have the predicted biological activities. The anti-viral activity of rhIFN-λ1 was determined as 10(6) IU/mg using the vesicular stomatitis virus (WISH-VSV) assay system. The anti-proliferation activity of rhIFN-λ1 was measured using the MTS method and the growth inhibition ratio was 57% higher than that for recombinant human IFN-α2b (rhIFN-α2b) when the rhIFN-λ1 concentration was 1000 IU/ml. rhIFN-λ1 had lower natural killer cell cytotoxicity than rhIFN-α2b. CONCLUSION: The Flp-In-CHO system is suitable for stably expressing rhIFN-λ1 that possesses the predicted anti-viral, anti-proliferation and natural killer cell cytotoxicity-promoting activities.
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spelling pubmed-57099782017-12-06 IFN-λ1 in CHO cells: its expression and biological activity Yuan, Wu-mei Zhang, Wan-ju Ma, Fen-lian Li, Jin-song Zhang, Qian Zheng, Li-shu Cell Mol Biol Lett Short Report BACKGROUND: Many studies have investigated the characteristics and biological activities of type III interferon (IFN), finding that it has similar features to type I IFN but also unique actions because it is recognized by a different receptor. RESULTS: A full-length recombinant human IFN-λ1 (rhIFN-λ1) cDNA was cloned into the pDF expression vector and stably expressed in Flp-In-CHO cells. After four purification steps (ammonium sulfate precipitation, SP Sepharose chromatography, Blue Sepharose 6 fast flow affinity chromatography and molecular sieve chromatography), the rhIFN-λ1 had a purity of about 90% and was found to have the predicted biological activities. The anti-viral activity of rhIFN-λ1 was determined as 10(6) IU/mg using the vesicular stomatitis virus (WISH-VSV) assay system. The anti-proliferation activity of rhIFN-λ1 was measured using the MTS method and the growth inhibition ratio was 57% higher than that for recombinant human IFN-α2b (rhIFN-α2b) when the rhIFN-λ1 concentration was 1000 IU/ml. rhIFN-λ1 had lower natural killer cell cytotoxicity than rhIFN-α2b. CONCLUSION: The Flp-In-CHO system is suitable for stably expressing rhIFN-λ1 that possesses the predicted anti-viral, anti-proliferation and natural killer cell cytotoxicity-promoting activities. BioMed Central 2017-12-01 /pmc/articles/PMC5709978/ /pubmed/29213292 http://dx.doi.org/10.1186/s11658-017-0057-x Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Short Report
Yuan, Wu-mei
Zhang, Wan-ju
Ma, Fen-lian
Li, Jin-song
Zhang, Qian
Zheng, Li-shu
IFN-λ1 in CHO cells: its expression and biological activity
title IFN-λ1 in CHO cells: its expression and biological activity
title_full IFN-λ1 in CHO cells: its expression and biological activity
title_fullStr IFN-λ1 in CHO cells: its expression and biological activity
title_full_unstemmed IFN-λ1 in CHO cells: its expression and biological activity
title_short IFN-λ1 in CHO cells: its expression and biological activity
title_sort ifn-λ1 in cho cells: its expression and biological activity
topic Short Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5709978/
https://www.ncbi.nlm.nih.gov/pubmed/29213292
http://dx.doi.org/10.1186/s11658-017-0057-x
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