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Aberrant localization and underglycosylation of highly accumulating single-chain Fv-Fc antibodies in transgenic Arabidopsis seeds. / Van Droogenbroeck, Bart; Cao, Jingyuan; Stadlmann, Johannes; Altmann, Friedrich; Colanesi, Sarah; Hillmer, Stefan; Robinson, David G; Van Lerberge, Els; Terryn, Nancy; Van Montagu, Marc; Liang, Mifang; Depicker, Ann; De Jaeger, Geert.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, Nr. 4, 2007, blz. 1430-5.

Onderzoeksoutput: Bijdrage aan tijdschriftA1: Web of Science-artikel

Harvard

Van Droogenbroeck, B, Cao, J, Stadlmann, J, Altmann, F, Colanesi, S, Hillmer, S, Robinson, DG, Van Lerberge, E, Terryn, N, Van Montagu, M, Liang, M, Depicker, A & De Jaeger, G 2007, 'Aberrant localization and underglycosylation of highly accumulating single-chain Fv-Fc antibodies in transgenic Arabidopsis seeds', Proceedings of the National Academy of Sciences of the United States of America, vol. 104, nr. 4, blz. 1430-5. https://doi.org/10.1073/pnas.0609997104

APA

Van Droogenbroeck, B., Cao, J., Stadlmann, J., Altmann, F., Colanesi, S., Hillmer, S., ... De Jaeger, G. (2007). Aberrant localization and underglycosylation of highly accumulating single-chain Fv-Fc antibodies in transgenic Arabidopsis seeds. Proceedings of the National Academy of Sciences of the United States of America, 104(4), 1430-5. https://doi.org/10.1073/pnas.0609997104

Vancouver

Van Droogenbroeck B, Cao J, Stadlmann J, Altmann F, Colanesi S, Hillmer S et al. Aberrant localization and underglycosylation of highly accumulating single-chain Fv-Fc antibodies in transgenic Arabidopsis seeds. Proceedings of the National Academy of Sciences of the United States of America. 2007;104(4):1430-5. https://doi.org/10.1073/pnas.0609997104

Author

Van Droogenbroeck, Bart ; Cao, Jingyuan ; Stadlmann, Johannes ; Altmann, Friedrich ; Colanesi, Sarah ; Hillmer, Stefan ; Robinson, David G ; Van Lerberge, Els ; Terryn, Nancy ; Van Montagu, Marc ; Liang, Mifang ; Depicker, Ann ; De Jaeger, Geert. / Aberrant localization and underglycosylation of highly accumulating single-chain Fv-Fc antibodies in transgenic Arabidopsis seeds. In: Proceedings of the National Academy of Sciences of the United States of America. 2007 ; Vol. 104, Nr. 4. blz. 1430-5.

Bibtex

@article{01d33d39fb1144e2a5adde226a888161,
title = "Aberrant localization and underglycosylation of highly accumulating single-chain Fv-Fc antibodies in transgenic Arabidopsis seeds",
abstract = "Production of high-value recombinant proteins in transgenic seeds is an attractive and economically feasible alternative to conventional systems based on mammalian cells and bacteria. In contrast to leaves, seeds allow high-level accumulation of recombinant proteins in a relatively small volume and a stable environment. We demonstrate that single-chain variable fragment (scFv)-Fc antibodies, with N-terminal signal sequence and C-terminal KDEL tag, can accumulate to very high levels as bivalent IgG-like antibodies in Arabidopsis thaliana seeds and illustrate that a plant-produced anti-hepatitis A virus scFv-Fc has similar antigen-binding and in vitro neutralizing activities as the corresponding full-length IgG. As expected, most scFv-Fc produced in seeds contained only oligomannose-type N-glycans, but, unexpectedly, 35-40{\%} was never glycosylated. A portion of the scFv-Fc was found in endoplasmic reticulum (ER)-derived compartments delimited by ribosome-associated membranes. Additionally, consistent with the glycosylation data, large amounts of the recombinant protein were deposited in the periplasmic space, implying a direct transport from the ER to the periplasmic space between the plasma membrane and the cell wall. Aberrant localization of the ER chaperones calreticulin and binding protein (BiP) and the endogenous seed storage protein cruciferin in the periplasmic space suggests that overproduction of recombinant scFv-Fc disturbs normal ER retention and protein-sorting mechanisms in the secretory pathway.",
keywords = "Arabidopsis, Electrophoresis, Polyacrylamide Gel, Glycosylation, Mass Spectrometry, Microscopy, Electron, Peptide Mapping, Plantibodies, Plants, Genetically Modified, Trypsin",
author = "{Van Droogenbroeck}, Bart and Jingyuan Cao and Johannes Stadlmann and Friedrich Altmann and Sarah Colanesi and Stefan Hillmer and Robinson, {David G} and {Van Lerberge}, Els and Nancy Terryn and {Van Montagu}, Marc and Mifang Liang and Ann Depicker and {De Jaeger}, Geert",
year = "2007",
doi = "10.1073/pnas.0609997104",
language = "English",
volume = "104",
pages = "1430--5",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "4",

}

RIS

TY - JOUR

T1 - Aberrant localization and underglycosylation of highly accumulating single-chain Fv-Fc antibodies in transgenic Arabidopsis seeds

AU - Van Droogenbroeck, Bart

AU - Cao, Jingyuan

AU - Stadlmann, Johannes

AU - Altmann, Friedrich

AU - Colanesi, Sarah

AU - Hillmer, Stefan

AU - Robinson, David G

AU - Van Lerberge, Els

AU - Terryn, Nancy

AU - Van Montagu, Marc

AU - Liang, Mifang

AU - Depicker, Ann

AU - De Jaeger, Geert

PY - 2007

Y1 - 2007

N2 - Production of high-value recombinant proteins in transgenic seeds is an attractive and economically feasible alternative to conventional systems based on mammalian cells and bacteria. In contrast to leaves, seeds allow high-level accumulation of recombinant proteins in a relatively small volume and a stable environment. We demonstrate that single-chain variable fragment (scFv)-Fc antibodies, with N-terminal signal sequence and C-terminal KDEL tag, can accumulate to very high levels as bivalent IgG-like antibodies in Arabidopsis thaliana seeds and illustrate that a plant-produced anti-hepatitis A virus scFv-Fc has similar antigen-binding and in vitro neutralizing activities as the corresponding full-length IgG. As expected, most scFv-Fc produced in seeds contained only oligomannose-type N-glycans, but, unexpectedly, 35-40% was never glycosylated. A portion of the scFv-Fc was found in endoplasmic reticulum (ER)-derived compartments delimited by ribosome-associated membranes. Additionally, consistent with the glycosylation data, large amounts of the recombinant protein were deposited in the periplasmic space, implying a direct transport from the ER to the periplasmic space between the plasma membrane and the cell wall. Aberrant localization of the ER chaperones calreticulin and binding protein (BiP) and the endogenous seed storage protein cruciferin in the periplasmic space suggests that overproduction of recombinant scFv-Fc disturbs normal ER retention and protein-sorting mechanisms in the secretory pathway.

AB - Production of high-value recombinant proteins in transgenic seeds is an attractive and economically feasible alternative to conventional systems based on mammalian cells and bacteria. In contrast to leaves, seeds allow high-level accumulation of recombinant proteins in a relatively small volume and a stable environment. We demonstrate that single-chain variable fragment (scFv)-Fc antibodies, with N-terminal signal sequence and C-terminal KDEL tag, can accumulate to very high levels as bivalent IgG-like antibodies in Arabidopsis thaliana seeds and illustrate that a plant-produced anti-hepatitis A virus scFv-Fc has similar antigen-binding and in vitro neutralizing activities as the corresponding full-length IgG. As expected, most scFv-Fc produced in seeds contained only oligomannose-type N-glycans, but, unexpectedly, 35-40% was never glycosylated. A portion of the scFv-Fc was found in endoplasmic reticulum (ER)-derived compartments delimited by ribosome-associated membranes. Additionally, consistent with the glycosylation data, large amounts of the recombinant protein were deposited in the periplasmic space, implying a direct transport from the ER to the periplasmic space between the plasma membrane and the cell wall. Aberrant localization of the ER chaperones calreticulin and binding protein (BiP) and the endogenous seed storage protein cruciferin in the periplasmic space suggests that overproduction of recombinant scFv-Fc disturbs normal ER retention and protein-sorting mechanisms in the secretory pathway.

KW - Arabidopsis

KW - Electrophoresis, Polyacrylamide Gel

KW - Glycosylation

KW - Mass Spectrometry

KW - Microscopy, Electron

KW - Peptide Mapping

KW - Plantibodies

KW - Plants, Genetically Modified

KW - Trypsin

U2 - 10.1073/pnas.0609997104

DO - 10.1073/pnas.0609997104

M3 - A1: Web of Science-article

C2 - 17227846

VL - 104

SP - 1430

EP - 1435

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 4

ER -