A fast and flexible PEG-mediated transient expression system in plants for high level expression of secreted recombinant proteins.
Plant expression systems offer a valuable alternative to traditional systems for the production of recombinant biopharmaceuticals. A highly efficient polyethyleneglycol (PEG)-mediated transient expression system for secreted recombinant proteins in plants has been developed. The human vascular endothelial growth factor 121 (rhVEGF) has been successfully expressed and efficiently secreted into the culture medium by transiently transformed moss protoplasts. In order to obtain secretion efficiency data, different expressed signal peptides were analysed and time course studies were performed with expression constructs containing different promoters. The transformation procedure was optimised for high level expression (up to 10 microg/ml) and successfully performed even with a transgenic glyco-engineered strain lacking plant-specific immunogenic sugar residues in N-glycans. The amount of rhVEGF was produced in such quantity that it allowed for the analysis of biological activity, silver-staining and Western blotting, revealing the correct formation and processing of the homodimer. This fast and flexible transient expression system enables feasibility studies and construct optimisation to be concluded within a few days, thus avoiding the time consuming step of having to generate stably transformed lines.
Baur
Armin
A
greenovation Biotech GmbH, Boetzinger Strasse 29b, 79111 Freiburg, Germany.
Kaufmann
Franz
F
Rolli
Helene
H
Weise
Andreas
A
Luethje
Rasmus
R
Berg
Birgit
B
Braun
Michael
M
Baeumer
Wolfgang
W
Kietzmann
Manfred
M
Reski
Ralf
R
Gorr
Gilbert
G
D019068:Bryopsida / Q000235:genetics* / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000378:metabolism*; D011092:Polyethylene Glycols; D015202:Protein Engineering / Q000379:methods*; D011523:Protoplasts / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection / Q000379:methods; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism*
D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D011092:Polyethylene Glycols