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| Hypoallergenic Derivatives of the Major Peanut Allergen Ara h 2 Obtained by Rational Sequence Reassembly |
| YI Hai-tao1,2, LIU Fang3, XIA Li-xin2,4, YAN Hao1,2, LIU Fei-yang2,4, LI Jian-jie1,2, LIU Zhi-gang2,4 |
1. College of Life Science,Shenzhen University, Shenzhen 518060, China;
2. Institute of Allergy and Immunology, Shenzhen University, Shenzhen 518060, China;
3. Xinjiang Huashidan Pharmaceutical Co., Ltd,Wulumuqi 830000,China;
4. Medical College, Shenzhen University, Shenzhen 518060, China |
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Abstract Objective: To express and purify the peanut major allergen Ara h 2 that obtained by rational sequence reassembly and preliminarily characterize the hypoallergenic of purified recombinant S-Ara h 2 protein. Methods: The Ara h 2 sequence 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 S-Ara h 2 protein. The hypoallergenic of S-Ara h 2 was examined by Western blotting and ELISA. Results: The ORF which contained 471 bp and encoded 157 amino acids was authenticated to be S-Ara h 2. The recombinant S-Ara h 2 protein,induced by IPTG,which is consistent with the actual value. The affinity between recombinant S-Ara h 2 protein and IgE antibodies from pooled peanut-allergic patients serum was significant decrease compared with R-Ara h 2 was identified by Western blotting and ELISA. Conclusion: Recombinant S-Ara h 2 protein was obtained with hypoallergenic. It will provide a novel safe vaccine to cure peanut allergy patients by Allergen-specific immunotherapy.
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Received: 31 March 2011
Published: 25 July 2011
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