Advanced search
1 file | 673.84 KB

Comparison of VHH-Fc antibody production in Arabidopsis thaliana, Nicotiana benthamiana and Pichia pastoris

Thomas De Meyer (UGent) , Bram Laukens (UGent) , Jonah Nolf (UGent) , Els Van Lerberge (UGent) , Riet De Rycke (UGent) , Ans De Beuckelaer (UGent) , Sylvie De Buck (UGent) , Nico Callewaert (UGent) and Anna Depicker (UGent)
(2015) PLANT BIOTECHNOLOGY JOURNAL. 13(7). p.938-947
Author
Organization
Abstract
VHHs or nanobodies are widely acknowledged as interesting diagnostic and therapeutic tools. However, for some applications, multivalent antibody formats, such as the dimeric VHH-Fc format, are desired to increase the functional affinity. The scope of this study was to compare transient expression of diagnostic VHH-Fc antibodies in Nicotiana benthamiana leaves with their stable expression in Arabidopsis thaliana seeds and Pichia pastoris. To this end, VHH-Fc antibodies targeting green fluorescent protein or the A.thaliana seed storage proteins (albumin and globulin) were produced in the three platforms. Differences were mainly observed in the accumulation levels and glycosylation patterns. Interestingly, although in plants oligomannosidic N-glycans were expected for KDEL-tagged VHH-Fcs, several VHH-Fcs with an intact KDEL-tag carried complex-type N-glycans, suggesting a dysfunctional retention in the endoplasmic reticulum. All VHH-Fcs were equally functional across expression platforms and several outperformed their corresponding VHH in terms of sensitivity in ELISA.
Keywords
RECOMBINANT ANTIBODIES, TRANSIENT EXPRESSION, PHARMACEUTICAL PROTEINS, MONOCLONAL-ANTIBODY, N-GLYCANS, PROTEIN-PRODUCTION, SINGLE-DOMAIN ANTIBODIES, avidity, N-glycosylation, recombinant protein production, molecular farming, heavy-chain antibody, nanobody, THERMAL-STABILITY, LINKED GLYCANS, IN-VIVO

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 673.84 KB

Citation

Please use this url to cite or link to this publication:

Chicago
De Meyer, Thomas, Bram Laukens, Jonah Nolf, Els Van Lerberge, Riet De Rycke, Ans De Beuckelaer, Sylvie De Buck, Nico Callewaert, and Anna Depicker. 2015. “Comparison of VHH-Fc Antibody Production in Arabidopsis Thaliana, Nicotiana Benthamiana and Pichia Pastoris.” Plant Biotechnology Journal 13 (7): 938–947.
APA
De Meyer, Thomas, Laukens, B., Nolf, J., Van Lerberge, E., De Rycke, R., De Beuckelaer, A., De Buck, S., et al. (2015). Comparison of VHH-Fc antibody production in Arabidopsis thaliana, Nicotiana benthamiana and Pichia pastoris. PLANT BIOTECHNOLOGY JOURNAL, 13(7), 938–947.
Vancouver
1.
De Meyer T, Laukens B, Nolf J, Van Lerberge E, De Rycke R, De Beuckelaer A, et al. Comparison of VHH-Fc antibody production in Arabidopsis thaliana, Nicotiana benthamiana and Pichia pastoris. PLANT BIOTECHNOLOGY JOURNAL. 2015;13(7):938–47.
MLA
De Meyer, Thomas et al. “Comparison of VHH-Fc Antibody Production in Arabidopsis Thaliana, Nicotiana Benthamiana and Pichia Pastoris.” PLANT BIOTECHNOLOGY JOURNAL 13.7 (2015): 938–947. Print.
@article{6951243,
  abstract     = {VHHs or nanobodies are widely acknowledged as interesting diagnostic and therapeutic tools. However, for some applications, multivalent antibody formats, such as the dimeric VHH-Fc format, are desired to increase the functional affinity. The scope of this study was to compare transient expression of diagnostic VHH-Fc antibodies in Nicotiana benthamiana leaves with their stable expression in Arabidopsis thaliana seeds and Pichia pastoris. To this end, VHH-Fc antibodies targeting green fluorescent protein or the A.thaliana seed storage proteins (albumin and globulin) were produced in the three platforms. Differences were mainly observed in the accumulation levels and glycosylation patterns. Interestingly, although in plants oligomannosidic N-glycans were expected for KDEL-tagged VHH-Fcs, several VHH-Fcs with an intact KDEL-tag carried complex-type N-glycans, suggesting a dysfunctional retention in the endoplasmic reticulum. All VHH-Fcs were equally functional across expression platforms and several outperformed their corresponding VHH in terms of sensitivity in ELISA.},
  author       = {De Meyer, Thomas and Laukens, Bram and Nolf, Jonah and Van Lerberge, Els and De Rycke, Riet and De Beuckelaer, Ans and De Buck, Sylvie and Callewaert, Nico and Depicker, Anna},
  issn         = {1467-7644},
  journal      = {PLANT BIOTECHNOLOGY JOURNAL},
  keywords     = {RECOMBINANT ANTIBODIES,TRANSIENT EXPRESSION,PHARMACEUTICAL PROTEINS,MONOCLONAL-ANTIBODY,N-GLYCANS,PROTEIN-PRODUCTION,SINGLE-DOMAIN ANTIBODIES,avidity,N-glycosylation,recombinant protein production,molecular farming,heavy-chain antibody,nanobody,THERMAL-STABILITY,LINKED GLYCANS,IN-VIVO},
  language     = {eng},
  number       = {7},
  pages        = {938--947},
  title        = {Comparison of VHH-Fc antibody production in Arabidopsis thaliana, Nicotiana benthamiana and Pichia pastoris},
  url          = {http://dx.doi.org/10.1111/pbi.12330},
  volume       = {13},
  year         = {2015},
}

Altmetric
View in Altmetric
Web of Science
Times cited: