Preparation and hypoallergenic characterization of Ara h 2 Trimer

This study is aimed to express and purify the peanut major allergen Ara h 2 Trimer and preliminarily characterize the hypoallergenic of purified recombinant trimer protein.Three Ara h 2 molecules were connected and inserted into the expression vector pET-32a(+).Then the vector was transformed into Origami and the protein expression was induced by IPTG.The recombinant trimer protein was purified by Ni2+ chelating affinity chromatography.And the hypoallergenic of Ara h 2 trimer protein was examined by Western-blotting and ELISA.The ORF containing 1434 bp and encoding 477 amino acids was authenticated to be Ara h 2 Trimer.The recombinant Ara h 2 Trimer protein,induced by IPTG,is consistent with the actual value.Western-blotting and ELISA pointed that the affinity of recombinant Ara h 2 trimer protein to IgE antibodies from pooled peanut-allergic patients serum was significant descended as compared with r-Ara h.All the results indicate that Recombinant Ara h 2 Trimer protein with hypoallergenic is obtained in this study.