| Abstract Detail
Plant biotech & Risk Assessment Beuf, Laurent [1], Olagnier, Béatrice [2], Laparra, Hélène [3], Comeau, David [4], Peyrot, Corinne [3], Montandon, Frédéric [5], Desmaizières, Gregory [5], Duchateau, Nathalie [2], Decombas, Patrice [3], van Ree, Ronald [6], Moingeon, Philippe [7], Burtin, Daniel [8]. Production of the house dust mite major allergen Der p 1 in tobacco plants. Der p 1 is one of the major allergen of house dust mite D. pteronyssinus, it is described in around 80% of the sensitised patients. Therapeutic vaccines against house dust mite allergy are currently based on complex biological extracts. Recombinant allergen production in transgenic plants would offer a safe and consistent alternative to the dust mite extracts. In this study different strategies were developed to express the recombinant Der p 1 into stably transformed tobacco plants. The Der p 1 protein belongs to the cystein protease family it is produced following maturation of a precursor by cleavage of a propeptide. N-glycosylation and proteolytic activity have been hypothesised as important factors for the maturation of the allergen. In order to assess the influence of glycosylation, propeptide requirement, and active site mutation on protein maturation, the Der p 1 coding sequence was modified to test all the different modifications individually or in combination. Der p 1 was detected in each individual leave of the transgenic tobacco plants, demonstrating the good stability of the recombinant Der p 1 during plant maturation. The recombinant allergen was quantified by ELISA using anti Der p 1 monoclonal antibodies, purified and characterized by biochemical and immunological methods. This study showed the feasibility to express at high level a complex glycosylated molecule with a propeptide, such as Der p 1 in transgenic tobacco plants. Various recombinant Der p1 forms produced were found to exhibit a similar immunoreactivity profile to the natural Der p1. The plant-based expression system developed by MERISTEM Therapeutics is an efficient and cost-effective tool to produce complex recombinant allergens in a native configuration for therapeutic or diagnostic purposes.
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1 - Meristem Therapeutics, Molecular Biology, 8 Rue des frères Lumière, Clermont-Ferrand, 63100, France
2 - Meristem Therapeutics, Biochemistry
3 - Meristem Therapeutics, Cell Biology
4 - Meristem Therapeutics, Molecular Biology
5 - Meristem Therapeutics, Purification
6 - Sanquin
7 - Stallergenes
8 - Meristem Therapeutics
Keywords:
Biotechnology
recombinant protein
allergen.
Presentation Type: Plant Biology Abstract
Session: P
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM
Number: P45017
Abstract ID:1244 |
