| High-level production of human interleukin-10 fusions in tobacco cell suspension cultures. | |
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MedLine Citation:
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PMID: 23297698 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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The production of pharmaceutical proteins in plants has made much progress in recent years with the development of transient expression systems, transplastomic technology and humanizing glycosylation patterns in plants. However, the first therapeutic proteins approved for administration to humans and animals were made in plant cell suspensions for reasons of containment, rapid scale-up and lack of toxic contaminants. In this study, we have investigated the production of human interleukin-10 (IL-10) in tobacco BY-2 cell suspension and evaluated the effect of an elastin-like polypeptide tag (ELP) and a green fluorescent protein (GFP) tag on IL-10 accumulation. We report the highest accumulation levels of hIL-10 obtained with any stable plant expression system using the ELP fusion strategy. Although IL-10-ELP has cytokine activity, its activity is reduced compared to unfused IL-10, likely caused by interference of ELP with folding of IL-10. Green fluorescent protein has no effect on IL-10 accumulation, but examining the trafficking of IL-10-GFP over the cell culture cycle revealed fluorescence in the vacuole during the stationary phase of the culture growth cycle. Analysis of isolated vacuoles indicated that GFP alone is found in vacuoles, while the full-size fusion remains in the whole-cell extract. This indicates that GFP is cleaved off prior to its trafficking to the vacuole. On the other hand, IL-10-GFP-ELP remains mostly in the ER and accumulates to high levels. Protein bodies were observed at the end of the culture cycle and are thought to arise as a consequence of high levels of accumulation in the ER. |
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Authors:
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Angelo Kaldis; Adil Ahmad; Alexandra Reid; Brian McGarvey; Jim Brandle; Shengwu Ma; Anthony Jevnikar; Susanne E Kohalmi; Rima Menassa |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2013-1-9 |
Journal Detail:
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Title: Plant biotechnology journal Volume: - ISSN: 1467-7652 ISO Abbreviation: Plant Biotechnol. J. Publication Date: 2013 Jan |
Date Detail:
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Created Date: 2013-1-9 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101201889 Medline TA: Plant Biotechnol J Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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© Her Majesty the Queen in Right of Canada 2013 Reproduced with the permission of the Minister of Agriculture and Agri-Food Canada. |
Affiliation:
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Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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