Transient Recombinant Protein Production in Glycoengineered Nicotiana benthamiana Cell Suspension Culture
AbstractTransient recombinant protein production is a promising alternative to stable transgenic systems, particularly for emergency situations in which rapid production of novel therapeutics is needed. In plants, Agrobacterium tumefaciens can be used as a gene delivery vector for transient expression. A potential barrier for plant-based production of human therapeutics is that different glycosylation patterns are found on plant and mammalian proteins. Since glycosylation can affect the efficacy, safety and stability of a therapeutic protein, methods to control glycan structures and distributions in plant-based systems would be beneficial. In these studies, we performed Agrobacterium-mediated transient expression in glycoengineered plant cell suspension cultures. To reduce the presence of plant-specific glycans on the product, we generated and characterized cell suspension cultures from β-1,2-xylosyltransferase and α-1,3-fucosyltransferase knockdown Nicotiana benthamiana. An anthrax decoy fusion protein was transiently produced in these glycoengineered plant cell suspension cultures through co-culture with genetically engineered Agrobacterium. The mass ratio of Agrobacterium to plant cells used was shown to impact recombinant protein expression levels. N-glycosylation analysis on the anthrax decoy fusion protein produced in glycoengineered N. benthamiana showed a dramatic reduction in plant-specific N-glycans. Overall, the results presented here demonstrate the feasibility of a simple, rapid and scalable process for transient production of recombinant proteins without plant-specific glycans in a glycoengineered plant cell culture host. View Full-Text
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Sukenik, S.C.; Karuppanan, K.; Li, Q.; Lebrilla, C.B.; Nandi, S.; McDonald, K.A. Transient Recombinant Protein Production in Glycoengineered Nicotiana benthamiana Cell Suspension Culture. Int. J. Mol. Sci. 2018, 19, 1205.
Sukenik SC, Karuppanan K, Li Q, Lebrilla CB, Nandi S, McDonald KA. Transient Recombinant Protein Production in Glycoengineered Nicotiana benthamiana Cell Suspension Culture. International Journal of Molecular Sciences. 2018; 19(4):1205.Chicago/Turabian Style
Sukenik, Sara C.; Karuppanan, Kalimuthu; Li, Qiongyu; Lebrilla, Carlito B.; Nandi, Somen; McDonald, Karen A. 2018. "Transient Recombinant Protein Production in Glycoengineered Nicotiana benthamiana Cell Suspension Culture." Int. J. Mol. Sci. 19, no. 4: 1205.
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