The development, characterization, and demonstration of a novel strategy for purification of recombinant proteins expressed in plants.

摘要: Plants have attracted increasing attention as an expression platform for the production of pharmaceutical proteins due to its unlimited scalability and low cost potential. However, compared other systems, plants accumulate relatively levels foreign proteins, thus necessitating development efficient systems purification from plant tissues. We developed a novel strategy recombinant expressed in plants, based on genetic fusion soybean agglutinin (SBA), homotetrameric lectin that binds N-acetyl-D-galactosamine. Previously it was shown high purity SBA could be recovered with efficiency greater than 90% following one-step using N-acetyl-D-galactosamine-agar columns. constructed protein containing reporter green fluorescent (GFP) transiently N. benthamiana plants. achieved over 2.5% TSP accumulation leaves benthamiana. Confocal microscopic analysis demonstrated vivo activity fused GFP partner. Importantly, rSBA-GFP crude leaf extract ~90% yield via columns, purified able induce agglutination rabbit red blood cells. Combined this, tetrameric assembly western blotting. In addition, retained signal agglutinated cells, demonstrating feasibility discrimination cells bear ligand glycan their surface. This work validates effective affinity tag simple rapid genetically proteins.