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Critical Amino Acids Analysis of Epitopes in Pen a1

文献类型： 外文期刊

作者： Mou Hui 1 ; Gao Mei-Xu 1 ; Li Shu-Rong 2 ; Wang Zhi-Dong 1 ; Pan Jia-Rong 3 ; Zhao Jie 1 ; Zhi Yu-Xiang 4 ; Li Shu-Jin 1 ; Z 1 ;

作者机构： 1.Chinese Acad Agr Sci, Inst Agroprod Proc Sci & Technol, Key Lab Agroprod Proc, Minist Agr, Beijing 100193, Peoples R China

2.Beijing Vocat Coll Agr, Beijing 102442, Peoples R China

3.China Jiliang Univ, Coll Iife Sci, Hangzhou 310018, Zhejiang, Peoples R China

4.Beijing Union Med Coll Hosp, Dept Allergy, Beijing 100052, Peoples R China

关键词： Pen a1;Epitope;Mutated polypeptides;Critical amino acids

期刊名称：CHINESE JOURNAL OF ANALYTICAL CHEMISTRY （ 影响因子：1.134； 五年影响因子：0.909 ）

ISSN： 0253-3820

年卷期： 2014 年 42 卷 11 期

页码：

收录情况： SCI

摘要： A screening method for critical amino acids in Epitopes using anti-Epitope antibody was developed. The amino acids' frequency of occurrence in Epitopes of shrimp antigen Pen a1 was calculated using MEGA5 software and their conservative property of tropomyosin in SDAP database bank was analyzed using DNAMAN software. Potential critical amino acids based on these two methods were identified, namely Epitope 1 (K, E, N), Epitope 2 (K, L, E), Epitope 3 (E, R, D, L, Q), Epitope 5 (K, L, Q). The mutated polypeptides corresponding to these Epitopes in which these critical amino acids were substituted with alanine were synthesized. The IgG binding ability of these mutants was analyzed by competitive immunodot-blot method and indirect ELISA using Epitope antibody to screen the critical amino acids. Experimental results showed that the critical amino acids of the four Epitopes were as follows: Epitope 1 (Gln), Epitope 2 (Leu and Glu), Epitope 3 (Leu and Asp), Epitope 5 (Leu). The feasibility of this screening method was proved and it also offered a theoretical foundation for research on sensitization mechanism of Pen a1 and desensitization using gene modification.

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