Analysis of the IgE epitope profile of soybean allergen Gly m 4
نویسندگان
چکیده
Methods Sera from 33 birch pollen-allergic patients with and without clinical reactivity to soy as determined by oral food challenge or convincing history of soy allergy were included in the study. Specific IgE against Bet v 1 and Gly m 4 was determined by ImmunoCAP. IgE binding of the sera to rGly m 4 and soy extract was tested in western blot analysis. To predict putative IgE epitopes, 20 IgE-binding phagedisplayed peptide mimics and heptamers representing the total Gly m 4 amino acid sequence were bioinformatically mapped onto the molecular surface of Gly m 4. To verify predicted epitopes, Gly m 4 variants with substituted epitope residues were expressed in and purified from E. coli. Secondary and tertiary structures were assessed by CD and NMR spectroscopy. IgE binding of variants was analyzed by competitive immunoblotting and inhibition ELISA.
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عنوان ژورنال:
دوره 4 شماره
صفحات -
تاریخ انتشار 2014