Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants
Chemical and Biomolecular Engineering(2018)
摘要
Apart from tradition, the biospecific affinity isolation has become one of the most rapidly growing concerns with cellulose binding domains (CBDs), a high-capacity tag for cost-effective purifications of fusion proteins. We report here a new strategy optimized for isolation of two fusion proteins, the FGF-1 (a human functional protein) and the H5N1 (a vaccine candidate antigen) tagged with CBD were grown in transient Nicotiana benthamiana and transgenic Arabidopsis thaliana respectively. A notable fraction of the recombinant proteins was lost through plant debris pelleted from the plant-slurry made. However, this issue was resolved by adjusting tissue-to-buffer ratios with 1:10 and 1:15 in those plants respectively. Washing efficiencies were improved by agitating column beds with acidic buffer (20mM NaAc. pH 4.0) in Nicotiana and alkaline buffer (10mM Tris-base pH 8.0) in Arabidopsis. Adsorption and coupling of tagged proteins on cellulose matrices were affected by the buffer-logged resins. The column-beds, after pumping the moisture out, showed efficient in binding of antigens with almost no losses detected by immunoblot signals. The bound antigens were released efficiently from the cellulose matrices by 1% Cellobiose and 2% Triethylamine respectively. The successive purifications of these antigenic proteins with identical tags likely indicate the efficiency of the proposed strategy in providing a generic and cost-effective method to purify fusion proteins propagated in transgenic plants.
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关键词
vaccine candidate antigens,cellulose-affinity
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