Designing and characterization of hypoallergenic derivative of Ara h 2,major allergen from peanut

The present study aims to express and purify a novel peanut major allergen Ara h 2 and analyze the hypoallergenic of purified recombinant T-Ara h 2 protein.The Ara h 2 was initiadly reassembled and then inserted into the expression vector pET-32a(+).The vector was transformed into Escherichia coli.Origami and the protein expression was induced by IPTG.Ni2+ chelating affinity chromatography was used to purify the recombinant T-Ara h 2 protein.The hypoallergenic of T-Ara h 2 was analyzed by Western-blotting and ELISA.The ORF(containing 453 bp and encoded 151 amino acids) was confirmed to be T-Ara h 2.The molecular weight recombinant T-Ara h 2 protein is consistent with the predicted value.The affinity(between recombinant T-Ara h 2 protein and IgE antibodies from pooled peanut-allergic patients serum) was significanly decrease compared with R-Ara h 2 in this process.Recombinant T-Ara h 2 protein was obtained with hypoallergenic.