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  • Efficient Agrobacterium-based transient expression system for the production of biopharmaceuticals in plants.

Efficient Agrobacterium-based transient expression system for the production of biopharmaceuticals in plants.

Bioengineered bugs (2011-02-18)
Patrizia Circelli, Marcello Donini, Maria Elena Villani, Eugenio Benvenuto, Carla Marusic
ABSTRACT

We have recently described an efficient transient expression system mediated by Agrobacterium tumefaciens for the production of HIV-1 Nef protein in Nicotiana benthamiana plants. In order to enhance the yield of recombinant protein we assayed the effect of three gene-silencing viral suppressor proteins (P25 of Potato Virus X, P19 of Artichoke Mottled Crinckle virus and Tomato Bushy Stunt virus) on Nef expression levels. Results demonstrated that AMCV-P19 gave the highest Nef yield (1.3% of total soluble protein) and that this effect was correlated to a remarkable decrease of Nef-specific small interfering RNAs (siRNAs) indicating an effective modulation of RNA silencing mechanisms. Here we report additional data on the production of different heterologous proteins including human immunoglobulin heavy and light chains and a virus coat protein that demonstrate the robustness of this co-agroinfiltration expression system boosted by the AMCV-P19 gene-silencing suppressor.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Human Lambda Light Chains (Bound and Free)−Peroxidase antibody produced in goat, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Monoclonal Anti-Human Lambda Light Chains (Bound and Free) antibody produced in mouse, clone HP-6054, ascites fluid
Sigma-Aldrich
Anti-Human IgG (γ-chain specific)−Peroxidase antibody produced in goat, IgG fraction of antiserum, buffered aqueous solution
Sigma-Aldrich
IgG1, Lambda from human myeloma plasma, purified immunoglobulin, >95% (SDS-PAGE)