Manduzio H.,ANGANY Genetics |
Fitchette A.-C.,ANGANY Genetics |
Hrabina M.,Stallergenes SA |
Chabre H.,Stallergenes SA |
And 5 more authors.
Plant Biotechnology Journal | Year: 2012
Grass pollen allergic patients are concomitantly exposed and sensitized to pollens from multiple Pooideae (i.e. common grass) species. As such, they are currently desensitized by allergen-specific immunotherapy using extracts made from mixes of pollens from Anthoxanthum odoratum, Dactylis glomerata, Lolium perenne, Phleum pratense and Poa pratensis. Herein, we demonstrate that species-specific glycoprotein patterns are documented by 1D and 2D electrophoresis and Western blotting analysis, which can be used as an identity test for such pollens. Most allergens are glycoproteins bearing complex N-glycans encompassing β1,2 xylose and α1,3 fucose glycoepitopes. Glycoepitope destruction using periodate oxidation has no impact on seric IgE reactivity in 75% atopic patients (n=24). The latter have thus no significant IgE responses to carbohydrate-containing epitopes. In contrast, periodate treatment strongly impairs IgE recognition of glycoallergens in 25% of patients tested, demonstrating the presence of carbohydrate-specific IgE in those patients. While the clinical impact of carbohydrate-specific IgE is still a matter of controversy, the presence of these IgE in the serum of many allergic patients illustrates the need for cross-reacting carbohydrate epitope-free recombinant allergens to develop relevant diagnostic tests. These data also support the pertinence of mixing multiple grass pollens to desensitize atopic patients, with the aim to broaden the repertoire of glycoepitopes in the vaccine, thus mimicking natural exposure conditions. © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.
Gomord V.,University of Rouen |
Fitchette A.,ANGANY Genetics |
Menu-Bouaouiche L.,University of Rouen |
Saint-Jore-Dupas C.,University of Rouen |
And 3 more authors.
Plant Biotechnology Journal | Year: 2010
While . N-glycan synthesis in the endoplasmic reticulum (ER) is relatively well conserved in eukaryotes, . N-glycan processing and . O-glycan biosynthesis in the Golgi apparatus are kingdom specific and result in different oligosaccharide structures attached to glycoproteins in plants and mammals. With the prospect of using plants as alternative hosts to mammalian cell lines for the production of therapeutic glycoproteins, significant progress has been made towards the humanization of protein . N-glycosylation in plant cells. To date, successful efforts in this direction have mainly focused on the targeted expression of therapeutic proteins, the knockout of plant-specific . N-glycan-processing genes, and/or the introduction of the enzymatic machinery catalyzing the synthesis, transport and addition of human sugars. By contrast, very little attention has been paid until now to the . O-glycosylation status of plant-made therapeutic proteins, which is surprising considering that hundreds of human proteins represent good candidates for Hyp-. O glycosylation when produced in a plant expression system. This review describes protein . N- and . O-linked glycosylation in plants and highlights the limitations and advantages of plant-specific glycosylation on plant-made biopharmaceuticals. © 2010 The Authors. Journal compilation © 2010 Blackwell Publishing Ltd.
Angany Genetics | Date: 2013-06-13
A method for producing a recombinant protein in a plant, in particular a tobacco plant, preferably Nicotiana benthamiana, includes the following steps: a) culturing the plant aeroponically or hydroponically, preferably on mobile floats and under LED lighting; b) vacuum agroinfiltration of the plant obtained in a) by agrobacteria that include a DNA fragment coding for the recombinant protein; c) returning the plants to culturing after step b), under the same conditions as for step a); and d) extracting and purifying the recombinant protein from the aerial portions of the plants produced in step c).