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Periodicum biologorum, Vol.118 No.2 Lipanj 2016.

Pregledni rad
https://doi.org/10.18054/pb.2016.118.2.3845

Recombinant therapeutic proteins produced in plants: towards engineering of human-type O-and N-glycosylation

Kruno Vukušić ; Laboratory of cell biophysics, Division of Molecular Biology, Ru|er Bo{kovi} Institute, Bijeni~ka cesta 54, HR-10000, Zagreb, Croatia
Sandra Šikić ; Department of Ecology, Dr. Andrija [tampar Institute of Public Health, Mirogojska cesta 16, HR-10000, Zagreb, Croatia
Biljana Balen ; Division of Molecular Biology, Faculty of Science, University of Zagreb, Horvatovac 102a, HR-10000, Zagreb, Croatia

Puni tekst: engleski, pdf (1 MB) str. 75-90 preuzimanja: 758* citiraj
APA 6th Edition
Vukušić, K., Šikić, S. i Balen, B. (2016). Recombinant therapeutic proteins produced in plants: towards engineering of human-type O-and N-glycosylation. Periodicum biologorum, 118 (2), 75-90. https://doi.org/10.18054/pb.2016.118.2.3845
MLA 8th Edition
Vukušić, Kruno, et al. "Recombinant therapeutic proteins produced in plants: towards engineering of human-type O-and N-glycosylation." Periodicum biologorum, vol. 118, br. 2, 2016, str. 75-90. https://doi.org/10.18054/pb.2016.118.2.3845. Citirano 16.12.2018.
Chicago 17th Edition
Vukušić, Kruno, Sandra Šikić i Biljana Balen. "Recombinant therapeutic proteins produced in plants: towards engineering of human-type O-and N-glycosylation." Periodicum biologorum 118, br. 2 (2016): 75-90. https://doi.org/10.18054/pb.2016.118.2.3845
Harvard
Vukušić, K., Šikić, S., i Balen, B. (2016). 'Recombinant therapeutic proteins produced in plants: towards engineering of human-type O-and N-glycosylation', Periodicum biologorum, 118(2), str. 75-90. doi: https://doi.org/10.18054/pb.2016.118.2.3845
Vancouver
Vukušić K, Šikić S, Balen B. Recombinant therapeutic proteins produced in plants: towards engineering of human-type O-and N-glycosylation. Periodicum biologorum [Internet]. 2016 [pristupljeno 16.12.2018.];118(2):75-90. doi: https://doi.org/10.18054/pb.2016.118.2.3845
IEEE
K. Vukušić, S. Šikić i B. Balen, "Recombinant therapeutic proteins produced in plants: towards engineering of human-type O-and N-glycosylation", Periodicum biologorum, vol.118, br. 2, str. 75-90, 2016. [Online]. doi: https://doi.org/10.18054/pb.2016.118.2.3845

Sažetak
Background and Purpose: Recombinant DNA technology has allowed expression of different heterologous proteins in many host systems, ranging from prokaryotic to eukaryotic organisms. Therapeutic properties of recombinant proteins are very often affected by the composition and heterogeneity of their glycans. Conventional expression systems for recombinant pharmaceuticals typically do not address this problem and result with products that contain a mixture of glycoforms that are neither identical to human glycans nor optimized for enhanced biological activity. Over the last decade plants have been developed as production platforms for recombinant proteins with pharmaceutical or industrial applications. Namely, plant expression systems contain very small differences in the post-translational modifications, mainly glycosylation, which can partly be overpowered by glycoengineering, whose goal is production of recombinant proteins with highly homogenous glycosylation that closely resembles the native system. This review attempts to present current accomplishments in the production of plant-derived glycoconjugates with humanized N- and O-glycans.

Materials and Methods: Main goal of N-glycoengineering is to reduce or eliminate plant-specific N-glycans, and at the same time to introduce mammalian-specific N-glycans through the several approaches. The easiest way is to change intracellular targeting of plant-made recombinant proteins and to ensure their retention in the ER; next approach is to eliminate the addition of plant-specific glycans; while the final step is engineering the plant glycosylation pathway to introduce mammalian glycotransferases into plants with generation of biantennary and multi-antennary structures on complex N-glycans. Due to significant differences in O-glycosylation between humans and plants, different approaches to engineering of O-glycosylation have been taken. Besides having their typical O-glycoslyation on Hyp-residues, plants in general miss the machinery for production of mammalian-type O-glycosylation. Attempts have been made to mimic mammalian O-glycosylation in plants, specifically the mucin-type addition of GalNAc residues.

Result: Efficient generation of bisected tetraantennary complex N-glycans without typical plant glycoepitopes on human erythropoietin (hEPO) and human transferrin (hTF) was obtained in Nicotiana benthamiana plants, thus demonstrating generation of recombinant proteins with human-type N-glycosylation at great uniformity. As for the O-glycosylation, attempts to produce mucin-type O-GalNAc and disialylated core 1 O-linked glycan structures on hEPO in N. benthamiana transgenic plants proved to be successful. Moreover, although small amounts of Hyp residues were found on recombinant EPO, no plant-specific O-glycans were detected, which demonstrates that plants are eligible candidates for production of recombinant therapeutics with fully humanized O- and N-glycans.

Conclusion: Plants and methods of plant molecular farming offer a powerful expression platform for the production of a variety of recombinant proteins, which show similar, or even higher, biological activity then protein or native homologs in cultured mammalian cells currently used for large-scale production.

Ključne riječi
plant molecular farming; glycoengineering; N-glycosylation; O-glycosylation

Hrčak ID: 162219

URI
https://hrcak.srce.hr/162219

Posjeta: 894 *