Products/Services Used | Details | Operation |
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Codon Optimization | cDNAs coding for hybrid molecules consisting of the major timothy grass pollen allergen Phl p 5a and the major birch pollen allergen Bet v 1a without linker (i.e., hybrid 1), with a flexible linker (3xGGGGS) (i.e., hybrid 2) or with a hydrophilic linker (3xSSSST) (i.e., hybrid 3) between the allergens and a 3′sequence coding for a hexahistidine tag were produced as synthetic genes with codons optimized for expression in E.coli and subcloned into the pET17b expression vector (GenScript, New Jersey, USA) (Fig. 1a). | Get A Quote |
The cross-linking of effector cell-bound IgE antibodies by allergens induces the release of inflammatory mediators which are responsible for the symptoms of allergy. We demonstrate that a recombinant hybrid molecule consisting of the major birch (Bet v 1) and grass (Phl p 5) pollen allergen exhibited reduced allergenic activity as compared to equimolar mixes of the isolated allergens in basophil activation experiments. The reduced allergenic activity of the hybrid was not due to reduced IgE reactivity as demonstrated by IgE binding experiments using sera from allergic patients. Physicochemical characterization of the hybrid by size exclusion chromatography, dynamic light scattering, negative-stain electron ... More