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Barley as a green factory for the production of functional Flt3 ligand
ORF Genetics, Keldnaholt, Reykjavík, Iceland.
Blood Systems Research Institute and Department of Laboratory Medicine, University of California San Francisco, San Francisco, CA, USA.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm , Ludwig Institute for Cancer Research.
ORF Genetics, Keldnaholt, Reykjavík, Iceland.
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2009 (English)In: Biotechnology Journal, ISSN 1860-7314, Vol. 5, no 2, 163-171 p.Article in journal (Refereed) Published
Abstract [en]

Biologically active recombinant human Flt3 ligand was expressed and isolated from transgenic barley seeds. Its expression is controlled by a tissue specific promoter that confines accumulation of the recombinant protein to the endosperm tissue of the seed. The recombinant Flt3 ligand variant expressed in the seeds contains an HQ-tag for affinity purification on immobilized metal ion affinity chromatography (IMAC) resin. The tagged protein was purified from seed extracts to near homogeneity using sequential chromatography on IMAC affinity resin and cation exchange resin. We also show that the recombinant Flt3 ligand protein undergoes posttranslational modifications: it is a glycoprotein containing alpha-1,3-fucose and alpha-1,2-xylose. The HQ-tagged Flt3 ligand variant exhibits comparable biological activity to commercial Flt3 ligand. This is the first report showing expression and accumulation of recombinant human growth factor in barley seeds with a yield of active protein similar to a bacterial expression system. The present results demonstrate that plant molecular farming is a viable approach for the bioproduction of human-derived growth factors.

Place, publisher, year, edition, pages
Weinheim: Wiley-VCH Verlag GmbH & Co. KGaA , 2009. Vol. 5, no 2, 163-171 p.
Keyword [en]
Animal-free, Expression system, Growth factors, Molecular farming, Recombinant human Flt3 ligand
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-114958DOI: 10.1002/biot.200900111ISI: 000275275500006PubMedID: 19844912OAI: oai:DiVA.org:uu-114958DiVA: diva2:295054
Available from: 2010-02-17 Created: 2010-02-17 Last updated: 2010-12-22Bibliographically approved

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