DOI: 10.1073/pnas.031419998. Galactose-extended glycans of antibodies produced by transgenic plants

Galactose-extended glycans of antibodies produced by transgenic plants

10.1073/pnas.031419998

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Plant-specificN-glycosylation can represent an important limitation for the use of recombinant glycoproteins of mammalian origin produced by transgenic plants. Comparison of plant and mammalianN-glycan biosynthesis indicates that β1,4-galactosyltransferase is the most important enzyme that is missing for conversion of typical plantN-glycans into mammalian-likeN-glycans. Here, the stable expression of human β1,4-galactosyltransferase in tobacco plants is described. Proteins isolated from transgenic tobacco plants expressing the mammalian enzyme bearN-glycans, of which about 15% exhibit terminal β1,4-galactose residues in addition to the specific plantN-glycan epitopes. The results indicate that the human enzyme is fully functional and localizes correctly in the Golgi apparatus. Despite the fact that through the modified glycosylation machinery numerous proteins have acquired unusualN-glycans with terminal β1,4-galactose residues, no obvious changes in the physiology of the transgenic plants are observed, and the feature is inheritable. The crossing of a tobacco plant expressing human β1,4-galactosyltransferase with a plant expressing the heavy and light chains of a mouse antibody results in the expression of a plantibody that exhibits partially galactosylatedN-glycans (30%), which is approximately as abundant as when the same antibody is produced by hybridoma cells. These results are a major step in thein plantaengineering of theN-glycosylation of recombinant antibodies.

Bibliography

Bakker, H., Bardor, M., Molthoff, J. W., Gomord, V., Elbers, I., Stevens, L. H., Jordi, W., Lommen, A., Faye, L., Lerouge, P., & Bosch, D. (2001). Galactose-extended glycans of antibodies produced by transgenic plants. Proceedings of the National Academy of Sciences, 98(5), 2899â2904.

Authors 11

Hans Bakker (first)
Muriel Bardor (additional)
Jos W. Molthoff (additional)
Véronique Gomord (additional)
Ingrid Elbers (additional)
Lucas H. Stevens (additional)
Wilco Jordi (additional)
Arjen Lommen (additional)
Loïc Faye (additional)
Patrice Lerouge (additional)
Dirk Bosch (additional)

References 36 Referenced 260

10.1016/S0167-7799(97)84202-3
10.1016/S0958-1669(99)80069-X
10.1038/nbt0896-975
10.1023/A:1006012005654
10.1074/jbc.275.15.11451
10.1093/glycob/9.4.365
10.1021/bi00512a001
10.1016/0161-5890(95)00118-2
10.1016/S0167-7799(96)10062-7
A Wright, S L Morrison J Immunol 160, 3393–3402 (1998). (10.4049/jimmunol.160.7.3393) / J Immunol by Wright A (1998)
10.1016/S0022-1759(98)00032-5
10.1038/nm0598-601
10.1016/S0006-291X(88)80300-0
10.1073/pnas.83.6.1573
10.1038/nbt1096-1288
10.1093/glycob/8.5.473
10.1073/pnas.96.8.4692
10.1093/emboj/16.8.1850
10.1073/pnas.96.3.974
10.1073/pnas.91.2.728
10.1073/pnas.89.10.4319
10.1074/jbc.270.20.12170
10.1007/BF01969123
10.1126/science.227.4691.1229
J Sambrook, E F Fritsch, T Maniatis Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Lab. Press, Plainview, NY, 1989). / Molecular Cloning: A Laboratory Manual by Sambrook J (1989)
10.1046/j.1365-313x.1997.12061411.x
10.1006/abio.1993.1088
10.1094/PHYTO.1997.87.8.839
10.1104/pp.121.2.333
10.1023/A:1006983125913
10.1002/(SICI)1098-2787(1999)18:6<349::AID-MAS1>3.0.CO;2-H
10.1046/j.1365-313X.1995.07010025.x
10.1104/pp.84.4.1301
10.1016/S0014-5793(97)01121-6
10.1093/glycob/10.5.477
10.1104/pp.102.4.1109

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:45 a.m.)
Deposited	2 years, 4 months ago (April 22, 2023, 8:37 a.m.)
Indexed	3 weeks, 6 days ago (Aug. 5, 2025, 8:23 a.m.)
Issued	24 years, 6 months ago (Feb. 27, 2001)
Published	24 years, 6 months ago (Feb. 27, 2001)
Published Online	24 years, 6 months ago (Feb. 27, 2001)
Published Print	24 years, 6 months ago (Feb. 27, 2001)

Funders 0

None

BibTeX

@article{Bakker_2001, title={Galactose-extended glycans of antibodies produced by transgenic plants}, volume={98}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.031419998}, DOI={10.1073/pnas.031419998}, number={5}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Bakker, Hans and Bardor, Muriel and Molthoff, Jos W. and Gomord, Véronique and Elbers, Ingrid and Stevens, Lucas H. and Jordi, Wilco and Lommen, Arjen and Faye, Loïc and Lerouge, Patrice and Bosch, Dirk}, year={2001}, month=feb, pages={2899–2904} }

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