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Int. J. Mol. Sci. 2018, 19(4), 1205; doi:10.3390/ijms19041205

Transient Recombinant Protein Production in Glycoengineered Nicotiana benthamiana Cell Suspension Culture

1
Department of Chemical Engineering, University of California, Davis, CA 95616, USA
2
Department of Chemistry, University of California, Davis, CA 95616, USA
3
Department of Biochemistry and Molecular Medicine, University of California, Davis, CA 95616, USA
4
Foods for Health Institute, University of California, Davis, CA 95616, USA
5
Global HealthShare Initiative, University of California, Davis, CA 95616, USA
*
Author to whom correspondence should be addressed.
Received: 24 February 2018 / Revised: 6 April 2018 / Accepted: 9 April 2018 / Published: 16 April 2018
(This article belongs to the Special Issue Recombinant Proteins)
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Abstract

Transient 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
Keywords: Agrobacterium tumefaciens; plant cell culture; co-culture; recombinant protein expression; N-glycosylation Agrobacterium tumefaciens; plant cell culture; co-culture; recombinant protein expression; N-glycosylation
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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.

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