Glikosile ve in vivo deglikosile edilmiş anti-PA83 formlarının Nicotiana benthamiana bitkisinde mühendisliği, üretimi ve karakterizasyonu

Since B. anthracis has been used as a potential agent, it is crucial to be able to develop an anthrax vaccine that is safe, low-cost, high-effective, and long-term stability. Latest studies show that plant-based transient expression systems are a promising technology that allows safe, rapid and inexpensive production of recombinant proteins due to their high expression capacity. Monoclonal antibody treatment is used as one of the way for limiting the spread and effects of the pathogen. Most of the full-length antibodies are produced in mammalian cell culture, bacteria, yeast, insects, and plant expression systems that have potential to produce recombinant antibodies. Plant expression systems are one of the most effective systems to produce recombinant therapeutic mAbs that can provide passive protection against pathogens. Plant expression systems are beneficial in terms of large-scale production, cost-efficiency, scalability, and safety. The plant produced monoclonal anti-PA83 antibody, which is capable of neutralizing toxin against anthrax, is an important target to develop reliable, effective and cost-effective vaccine candidates against anthrax. In recent studies; anti-PA83 monoclonal antibody produced using plant expression system and effective against anthrax protective antigen was produced in plants glycosylated and deglycosylated by mutation (Mett et al. 2011). In this thesis, in vivo PNGase F (Mamedov et al. 2012) and Endo H (Mamedov et al. 2017) deglycosylation strategy was used for deglycolysis of anti-PA83 protein. In this study, glycosylated and deglycosylated forms of the full-length anti-PA83 proteins were produced by co-expressed with the HC and LC genes that were HC and LC genes which were codon optimized and transferred to Agrobacterium competent cells in previous studies. The produced proteins were used to examine the antibody-antigen interaction the protein stability and toxin neutralizing ability of antibodies. In results of thesis, Endo H deglycosylated anti-PA83 showed more stability and activity than other anti-PA83 variants and this indicates that Endo H deglysocylated anti-PA83 can be used as a good vaccine candidate.