Comparison of expression levels of deglycosylated PA83 antigen produced by different plant expression vectors.

In recent years, many groups are selecting plants as the production hosts for recombinant proteins. The reasons are obvious—plants are free of animal and human pathogens, provide eukaryotic type of posttranslational modifications, including N-linked glycosylation and offer virtually unlimited scalability for protein production. . These technology technical and safety aspects of plants as production hosts including scalability of plant-based production,purification costs, yield, engineering of desired glycosylation pattern, and other topics. The successful use of Bacillus anthracis as a lethal biological weapon has prompted renewed research interest in the development of more effective vaccines against anthrax. Plant based vaccines offer safe alternative for vaccine production. Protective antigen (PA) of B. anthracis is an important component of anthrax toxin but its not a glycoprotein.However, its sequence has nine potential glycosylation sites and it has been shown to be aberrantly glycosylated when expressed in Nicotiana benthamiana plants. Recently, a non-glycosylated versions of PA83 antigen was produced by in vivo deglycosylation strategy (PNGase F or Endo H based) (Mamedov et al. 2012; Mamedov et al. 2016; Mamedov et al. 2017) or by site-directed mutagenesis-based approach, by mutation of six N-Glycosylations sites (Mamedov et al. 2016) In this study, new Anthrax vaccine candidates were identified by comparing the expression levels of the deglycosylated PA83 antigen produced by different plant expression vectors.a new mutant form by mutation of seven N-glycosylation sites (including Asn-39) of protective antigen was engineered, codon optimized, (Mamedov et al. 2017)expressed and produced by different plant expression vectors in N.benthamiana plant. Deglycosylated antigens of PA83 were purified using the IMAC affinity column chromatography method and evaluated for purified protein stability analysis.