Purification and site-specific N-glycosylation analysis of human recombinant butyrylcholinesterase from Nicotiana benthamiana

Effects of Kifunensine on Production and <em>N</em>-glycosylation Modification of Butyrylcholinesterase in a Transgenic Rice Cell Culture Bioreactor

“…In Arabidopsis thaliana, mutation of class I α-mannosidases (mns) genes resulted in swollen roots and altered cell wall structures in mutants [39]. It could be implied (Figure 3a, lane CA-) through image analysis, which is consistent with our previous reports using the same steps [2,3]. However, culture medium rrBChE in the absence of kifunensine was only 31% purity (Figure 3a with a single step Hupresin ® , while 99% purity of hBChE was achieved when Q-ceramic ion exchange chromatography at pH 4.5 was used prior the Hupresin ® step [42].…”

“…Protein expression in plant systems has the potential to provide a safe, cost-effective, and scalable method to meet the increasing need for therapeutic protein production. Plantbased expression offers several advantages to the biopharmaceutical industry, including decreased cost of production, scalability, a lack of susceptibility to mammalian pathogens, elimination of animal or human sourced raw materials, and the ability to produce complex proteins with post-translational modifications such as N-glycosylation [1][2][3][4][5]. For many therapeutic proteins, N-glycosylation is essential for protein folding, oligomerization, quality control, enzyme activity, ligand interactions, localization, and trafficking [6,7].…”

“…Even though TFF and DEAE were used for rrBChE purity is probably not high enough to prevent non-specific binding of non-target proteins to Hupresin ® . Alkanaimsh et al also found contaminating protein bands from Hupresin ® elution due to non-specific binding when non-enriched Nicotiana benthamiana rBChE cell extract was loaded onto Hupresin ®[2]. Starting with low BChE purity, Onder et al obtained 10-15% purity of hBChE purified from human blood plasma…”

Expression optimization, purification and in vitro characterization of human epidermal growth factor produced in Nicotiana benthamiana

“…N. benthamiana is a preferred host for the production of functional recombinant proteins such as cytokines, enzymes, therapeutic proteins, antibodies, and vaccines [ 15 , 19 ]. However, some of the limitations of plant-based platform includes low expression levels and difficulties in downstream processing/purification which have an immense effect on both cost and quality of the plant-produced proteins [ 20 ]. These limitations need to be addressed in plant expression platform in order to compete with the conventional expression system.…”

Effects of Kifunensine on Production and N-Glycosylation Modification of Butyrylcholinesterase in a Transgenic Rice Cell Culture Bioreactor

“…Protein expression in plant systems has the potential to provide a safe, cost-effective, and scalable method to meet the increasing need for therapeutic protein production. Plant-based expression offers several advantages to the biopharmaceutical industry, including decreased cost of production, scalability, lack of susceptibility to mammalian pathogens, elimination of animal- or human-sourced raw materials, and the production of complex proteins with post-translational modifications such as N -glycosylation [ 1 , 2 , 3 , 4 , 5 ]. For many therapeutic proteins, N -glycosylation is essential for protein folding, oligomerization, quality control, enzyme activity, ligand interactions, localization, and trafficking [ 6 , 7 ].…”