Ghent University Academic Bibliography

Advanced

Characterization of the single-chain Fv-Fc antibody MBP10 produced in Arabidopsis alg3 mutant seeds

Maurice Henquet, Jochem Eigenhuijsen, Thamara Hesselink, Holger Spiegel, Mariëlle Schreuder, Esther van Duijn, Jan Cordewener, Anna Depicker UGent, Alexander van der Krol and Dirk Bosch (2011) TRANSGENIC RESEARCH. 20(5). p.1033-1042
abstract
ER resident glycoproteins, including ectopically expressed recombinant glycoproteins, carry so-called high-mannose type N-glycans, which can be at different stages of processing. The presence of heterogeneous high-mannose type glycans on ER-retained therapeutic proteins is undesirable for specific therapeutic applications. Previously, we described an Arabidopsis alg3-2 glycosylation mutant in which aberrant Man(5)GlcNAc(2) mannose type N-glycans are transferred to proteins. Here we show that the alg3-2 mutation reduces the N-glycan heterogeneity on ER resident glycoproteins in seeds. We compared the properties of a scFv-Fc, with a KDEL ER retention tag (MBP10) that was expressed in seeds of wild type and alg3-2 plants. N-glycans on these antibodies from mutant seeds were predominantly of the intermediate Man5GlcNAc2 compared to Man(8)GlcNAc(2) and Man(7)GlcNAc(2) isoforms on MBP10 from wild-type seeds. The presence of aberrant N-glycans on MBP10 did not seem to affect MBP10 dimerisation nor binding of MBP10 to its antigen. In alg3-2 the fraction of underglycosylated MBP10 protein forms was higher than in wild type. Interestingly, the expression of MBP10 resulted also in underglycosylation of other, endogenous glycoproteins.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
Recombinant antibody, N-glycosylation, N-ACETYLGLUCOSAMINYLTRANSFERASE-I, Arabidopsis, Molecular farming, HUMAN BETA-1, 4-GALACTOSYLTRANSFERASE, MONOCLONAL-ANTIBODY, MASS-SPECTROMETRY, TRANSGENIC PLANTS, LINKED GLYCANS, GLYCOSYLATION, CELLS, PROTEIN
journal title
TRANSGENIC RESEARCH
Transgenic Res.
volume
20
issue
5
pages
1033 - 1042
Web of Science type
Article
Web of Science id
000296786900006
JCR category
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
JCR impact factor
2.754 (2011)
JCR rank
56/157 (2011)
JCR quartile
2 (2011)
ISSN
0962-8819
DOI
10.1007/s11248-010-9475-5
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
1982170
handle
http://hdl.handle.net/1854/LU-1982170
date created
2012-01-10 16:42:08
date last changed
2016-12-19 15:45:27
@article{1982170,
  abstract     = {ER resident glycoproteins, including ectopically expressed recombinant glycoproteins, carry so-called high-mannose type N-glycans, which can be at different stages of processing. The presence of heterogeneous high-mannose type glycans on ER-retained therapeutic proteins is undesirable for specific therapeutic applications. Previously, we described an Arabidopsis alg3-2 glycosylation mutant in which aberrant Man(5)GlcNAc(2) mannose type N-glycans are transferred to proteins. Here we show that the alg3-2 mutation reduces the N-glycan heterogeneity on ER resident glycoproteins in seeds. We compared the properties of a scFv-Fc, with a KDEL ER retention tag (MBP10) that was expressed in seeds of wild type and alg3-2 plants. N-glycans on these antibodies from mutant seeds were predominantly of the intermediate Man5GlcNAc2 compared to Man(8)GlcNAc(2) and Man(7)GlcNAc(2) isoforms on MBP10 from wild-type seeds. The presence of aberrant N-glycans on MBP10 did not seem to affect MBP10 dimerisation nor binding of MBP10 to its antigen. In alg3-2 the fraction of underglycosylated MBP10 protein forms was higher than in wild type. Interestingly, the expression of MBP10 resulted also in underglycosylation of other, endogenous glycoproteins.},
  author       = {Henquet, Maurice and Eigenhuijsen, Jochem and Hesselink, Thamara and Spiegel, Holger and Schreuder, Mari{\"e}lle and van Duijn, Esther and Cordewener, Jan and Depicker, Anna and van der Krol, Alexander and Bosch, Dirk},
  issn         = {0962-8819},
  journal      = {TRANSGENIC RESEARCH},
  keyword      = {Recombinant antibody,N-glycosylation,N-ACETYLGLUCOSAMINYLTRANSFERASE-I,Arabidopsis,Molecular farming,HUMAN BETA-1,4-GALACTOSYLTRANSFERASE,MONOCLONAL-ANTIBODY,MASS-SPECTROMETRY,TRANSGENIC PLANTS,LINKED GLYCANS,GLYCOSYLATION,CELLS,PROTEIN},
  language     = {eng},
  number       = {5},
  pages        = {1033--1042},
  title        = {Characterization of the single-chain Fv-Fc antibody MBP10 produced in Arabidopsis alg3 mutant seeds},
  url          = {http://dx.doi.org/10.1007/s11248-010-9475-5},
  volume       = {20},
  year         = {2011},
}

Chicago
Henquet, Maurice, Jochem Eigenhuijsen, Thamara Hesselink, Holger Spiegel, Mariëlle Schreuder, Esther van Duijn, Jan Cordewener, Anna Depicker, Alexander van der Krol, and Dirk Bosch. 2011. “Characterization of the Single-chain Fv-Fc Antibody MBP10 Produced in Arabidopsis Alg3 Mutant Seeds.” Transgenic Research 20 (5): 1033–1042.
APA
Henquet, M., Eigenhuijsen, J., Hesselink, T., Spiegel, H., Schreuder, M., van Duijn, E., Cordewener, J., et al. (2011). Characterization of the single-chain Fv-Fc antibody MBP10 produced in Arabidopsis alg3 mutant seeds. TRANSGENIC RESEARCH, 20(5), 1033–1042.
Vancouver
1.
Henquet M, Eigenhuijsen J, Hesselink T, Spiegel H, Schreuder M, van Duijn E, et al. Characterization of the single-chain Fv-Fc antibody MBP10 produced in Arabidopsis alg3 mutant seeds. TRANSGENIC RESEARCH. 2011;20(5):1033–42.
MLA
Henquet, Maurice, Jochem Eigenhuijsen, Thamara Hesselink, et al. “Characterization of the Single-chain Fv-Fc Antibody MBP10 Produced in Arabidopsis Alg3 Mutant Seeds.” TRANSGENIC RESEARCH 20.5 (2011): 1033–1042. Print.