Barley as a green factory for the production of functional Flt3 ligand.
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.
Erlendsson
Lýdur S
LS
ORF Genetics, Keldnaholt, Reykjavík, Iceland. lydur@orfgenetics.com
Muench
Marcus O
MO
Hellman
Ulf
U
Hrafnkelsdóttir
Soffía M
SM
Jonsson
Anders
A
Balmer
Yves
Y
Mäntylä
Einar
E
Orvar
Björn L
BL
D048708:Cell Growth Processes / Q000502:physiology; D045744:Cell Line, Tumor; D003001:Cloning, Molecular / Q000379:methods*; D005434:Flow Cytometry; D001467:Hordeum / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012639:Seeds / Q000235:genetics / Q000378:metabolism
D008565:Membrane Proteins; D011994:Recombinant Proteins; C084881:flt3 ligand protein