Current address: Istituto di Biologia e Biotecnologia Agraria, Consiglio Nazionale delle Ricerche, 20133 Milano, Italy.
Non-food/feed seeds as biofactories for the high-yield production of recombinant pharmaceuticals
Article first published online: 11 APR 2011
© 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd
Plant Biotechnology Journal
Volume 9, Issue 8, pages 911–921, October 2011
How to Cite
Morandini, F., Avesani, L., Bortesi, L., Van Droogenbroeck, B., De Wilde, K., Arcalis, E., Bazzoni, F., Santi, L., Brozzetti, A., Falorni, A., Stoger, E., Depicker, A. and Pezzotti, M. (2011), Non-food/feed seeds as biofactories for the high-yield production of recombinant pharmaceuticals. Plant Biotechnology Journal, 9: 911–921. doi: 10.1111/j.1467-7652.2011.00605.x
- Issue published online: 5 SEP 2011
- Article first published online: 11 APR 2011
- Received 6 September 2010; revised 8 February 2011; accepted 13 February 2011.
- molecular farming;
- plant made pharmaceuticals;
- transgenic plants;
- seed-specific expression;
- type 1 diabetes;
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® 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.