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Non-food/feed seeds as biofactories for the high-yield production of recombinant pharmaceuticals

Francesca Morandini, Linda Avesani, Luisa Bortesi, Bart Van Droogenbroeck UGent, Kirsten De Wilde, Elsa Arcalis, Flavia Bazzoni, Luca Santi, Annalisa Brozzetti, Alberto Falorni, et al. (2011) PLANT BIOTECHNOLOGY JOURNAL. 9(8). p.911-921
abstract
We describe an attractive cloning system for the seed-specific expression of recombinant proteins using three non-food/feed crops. A vector designed for direct subcloning by Gateway (R) recombination was developed and tested in Arabidopsis, tobacco and petunia plants for the production of a chimeric form (GAD67/65) of the 65 kDa isoform of glutamic acid decarboxylase (GAD65). GAD65 is one of the major human autoantigens involved in type 1 diabetes (T1D). The murine anti-inflammatory cytokine interleukin-10 (IL-10) was expressed with the described system in Arabidopsis and tobacco, whereas proinsulin, another T1D major autoantigen, was expressed in Arabidopsis. The cost-effective production of these proteins in plants could allow the development of T1D prevention strategies based on the induction of immunological tolerance. The best yields were achieved in Arabidopsis seeds, where GAD67/65 reached 7.7% of total soluble protein (TSP), the highest levels ever reported for this protein in plants. IL-10 and proinsulin reached 0.70% and 0.007% of TSP, respectively, consistent with levels previously reported in other plants or tissues. This versatile cloning vector could be suitable for the high-throughput evaluation of expression levels and stability of many valuable and difficult to produce proteins.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
transgenic plants, seed-specific expression, plant made pharmaceuticals, molecular farming, type 1 diabetes, GAD65, CHAIN VARIABLE FRAGMENTS, INSULIN FUSION PROTEIN, TOXIN-B-SUBUNIT, TRANSGENIC TOBACCO, ENDOPLASMIC-RETICULUM, ARABIDOPSIS-THALIANA, AUTOIMMUNE-DISEASES, GOLGI-APPARATUS, HUMAN CYTOMEGALOVIRUS, EXPRESSION SYSTEMS
journal title
PLANT BIOTECHNOLOGY JOURNAL
Plant Biotechnol. J.
volume
9
issue
8
pages
911 - 921
Web of Science type
Article
Web of Science id
000295042200010
JCR category
PLANT SCIENCES
JCR impact factor
5.442 (2011)
JCR rank
10/189 (2011)
JCR quartile
1 (2011)
ISSN
1467-7644
DOI
10.1111/j.1467-7652.2011.00605.x
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
1937439
handle
http://hdl.handle.net/1854/LU-1937439
date created
2011-10-27 16:17:48
date last changed
2016-12-19 15:42:08
@article{1937439,
  abstract     = {We describe an attractive cloning system for the seed-specific expression of recombinant proteins using three non-food/feed crops. A vector designed for direct subcloning by Gateway (R) recombination was developed and tested in Arabidopsis, tobacco and petunia plants for the production of a chimeric form (GAD67/65) of the 65 kDa isoform of glutamic acid decarboxylase (GAD65). GAD65 is one of the major human autoantigens involved in type 1 diabetes (T1D). The murine anti-inflammatory cytokine interleukin-10 (IL-10) was expressed with the described system in Arabidopsis and tobacco, whereas proinsulin, another T1D major autoantigen, was expressed in Arabidopsis. The cost-effective production of these proteins in plants could allow the development of T1D prevention strategies based on the induction of immunological tolerance. The best yields were achieved in Arabidopsis seeds, where GAD67/65 reached 7.7\% of total soluble protein (TSP), the highest levels ever reported for this protein in plants. IL-10 and proinsulin reached 0.70\% and 0.007\% of TSP, respectively, consistent with levels previously reported in other plants or tissues. This versatile cloning vector could be suitable for the high-throughput evaluation of expression levels and stability of many valuable and difficult to produce proteins.},
  author       = {Morandini, Francesca and Avesani, Linda and Bortesi, Luisa and Van Droogenbroeck, Bart and De Wilde, Kirsten and Arcalis, Elsa and Bazzoni, Flavia and Santi, Luca and Brozzetti, Annalisa and Falorni, Alberto and Stoger, Eva and Depicker, Anna and Pezzotti, Mario},
  issn         = {1467-7644},
  journal      = {PLANT BIOTECHNOLOGY JOURNAL},
  keyword      = {transgenic plants,seed-specific expression,plant made pharmaceuticals,molecular farming,type 1 diabetes,GAD65,CHAIN VARIABLE FRAGMENTS,INSULIN FUSION PROTEIN,TOXIN-B-SUBUNIT,TRANSGENIC TOBACCO,ENDOPLASMIC-RETICULUM,ARABIDOPSIS-THALIANA,AUTOIMMUNE-DISEASES,GOLGI-APPARATUS,HUMAN CYTOMEGALOVIRUS,EXPRESSION SYSTEMS},
  language     = {eng},
  number       = {8},
  pages        = {911--921},
  title        = {Non-food/feed seeds as biofactories for the high-yield production of recombinant pharmaceuticals},
  url          = {http://dx.doi.org/10.1111/j.1467-7652.2011.00605.x},
  volume       = {9},
  year         = {2011},
}

Chicago
Morandini, Francesca, Linda Avesani, Luisa Bortesi, Bart Van Droogenbroeck, Kirsten De Wilde, Elsa Arcalis, Flavia Bazzoni, et al. 2011. “Non-food/feed Seeds as Biofactories for the High-yield Production of Recombinant Pharmaceuticals.” Plant Biotechnology Journal 9 (8): 911–921.
APA
Morandini, F., Avesani, L., Bortesi, L., Van Droogenbroeck, B., De Wilde, K., Arcalis, E., Bazzoni, F., et al. (2011). Non-food/feed seeds as biofactories for the high-yield production of recombinant pharmaceuticals. PLANT BIOTECHNOLOGY JOURNAL, 9(8), 911–921.
Vancouver
1.
Morandini F, Avesani L, Bortesi L, Van Droogenbroeck B, De Wilde K, Arcalis E, et al. Non-food/feed seeds as biofactories for the high-yield production of recombinant pharmaceuticals. PLANT BIOTECHNOLOGY JOURNAL. 2011;9(8):911–21.
MLA
Morandini, Francesca, Linda Avesani, Luisa Bortesi, et al. “Non-food/feed Seeds as Biofactories for the High-yield Production of Recombinant Pharmaceuticals.” PLANT BIOTECHNOLOGY JOURNAL 9.8 (2011): 911–921. Print.