@entity56 to food are on the rise worldwide and there is a corresponding increase in interest to understand the molecular mechanisms responsible. @entity16244 are the most problematic because the reaction often persists into adulthood and can be as severe as @entity2021 and @entity204 . The purpose of the work presented here was to develop a reproducible method to produce large quantities of pure recombinant @entity8182 @entity7285 ( @entity8182 @entity7285 ) that will enable standardization of immunological tests for @entity1 and allow structural and immunological studies on the wild type and mutagenized forms of the protein. @entity8182 @entity7285 is initially a pre-pro-protein which, following two endoproteolytic cleavages, becomes the mature form found in peanut. The mature form however has flexible regions that make it refractory to some structural studies including crystallography. Therefore, independent purification of the mature and core regions was desirable. Expression constructs were synthesized cDNA clones for each in a pET plasmid vector without tags. Codons were optimized for expression in @entity526 . High-level expression was achieved in BL21 strains. Purification to near homogeneity was achieved by a combination of @entity1329 precipitation and ion exchange chromatography. The purified @entity8182 @entity7285 was then compared with natural @entity8182 @entity7285 for IgE binding. All @entity1 recognized both the folded natural and @entity8182 @entity7285 , but the IgE binding to the rArah1 was significantly reduced in comparison to the natural allergen, which could potentially make it useful for immunotherapeutic purposes.

Purification of Recombinant Peanut XXXX and Comparison of IgE Binding to the Natural Protein.

@entity1 :: ('9606', 'Species') :: ['patients'] @entity16244 :: ('MESH:D021183', 'Disease') :: ['Peanut allergies'] @entity526 :: ('562', 'Species') :: ['E. coli'] @entity7285 :: ('3009', 'Gene') :: ['h 1'] @entity56 :: ('MESH:D004342', 'Disease') :: ['Allergic reactions'] @entity1329 :: ('MESH:D000645', 'Chemical') :: ['ammonium sulfate'] @entity204 :: ('MESH:D003643', 'Disease') :: ['death'] @entity8182 :: ('24705(Tax:10116)', 'Gene') :: ['rAra', 'Ara'] @entity2021 :: ('MESH:D000707', 'Disease') :: ['anaphylaxis']
@entity7285 :: (3009,Gene) :: ['Allergen Ara h 1']
