Preparation and Characterization of the Genetically Engineered Hypoallergenic Derivative of the Major Peanut Allergen Ara h 2

The genetically engineered major allergen Ara h 2 was expressed and purified and the hypoallergenic of purified recombinant F-Ara h 2 protein was characterized.The Ara h 2 was reassembled and inserted into the expression vector pET-32a(+).The vector was transformed into Origami and the protein expression was induced by IPTG.Ni2+ Chelating affinity chromatography was used to purify the recombinant F-Ara h 2 protein.The hypoallergenic of F-Ara h 2 was examined by Western blotting and ELISA.The ORF which contained 471 bp and encoded 157 amino acids was authenticated to be F-Ara h 2.The recombinant F-Ara h 2 protein induced by IPTG was consistent with the actual value.The affinity between recombinant F-Ara h 2 protein and IgE antibodies from pooled peanut-allergic patients serum was significantly decreased compared with R-Ara h 2 was identified by Western blotting and ELISA.Recombinant F-Ara h 2 protein was hypoallergenic.