Identification of a Highly Conserved Epitope on Avian Influenza Virus Non-Structural Protein 1 Using a Peptide Microarray

文献类型： 外文期刊

第一作者： Sun, Jiashan

作者： Sun, Jiashan;Shi, Lin;Zheng, Shimin;Sun, Jiashan;Wang, Xiurong;Wen, Xuexia;Bao, Hongmei;Tao, Qimeng;Jiang, Yongping;Zeng, Xianying;Xu, Xiaolong;Tian, Guobin;Chen, Hualan;Sun, Jiashan;Wang, Xiurong;Wen, Xuexia;Bao, Hongmei;Tao, Qimeng;Jiang, Yongping;Zeng, Xianying;Xu, Xiaolong;Tian, Guobin;Chen, Hualan

作者机构：

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2016 年 11 卷 3 期

页码：

收录情况： SCI

摘要： Avian influenza virus (AIV) non-structural protein 1 (NS1) is a multifunctional protein. It is present at high levels in infected cells and can be used for AIV detection and diagnosis. In this study, we generated monoclonal antibody (MAb) D7 against AIV NS1 protein by immunization of BALB/c mice with purified recombinant NS1 protein expressed in Escherichia coli. Isotype determination revealed that the MAb was IgG(1)/kappa-type subclass. To identify the epitope of the MAb D7, the NS1 protein was truncated into a total of 225 15-mer peptides with 14 amino acid overlaps, which were spotted for a peptide microarray. The results revealed that the MAb D7 recognized the consensus DAPF motif. Furthermore, the AIV NS1 protein with the DAPF motif deletion was transiently expressed in 293T cells and failed to react with MAb D7. Subsequently, the DAPF motif was synthesized with an elongated GSGS linker at both the C-and N-termini. The MAb D7 reacted with the synthesized peptide both in enzyme-linked immunosorbent assay (ELISA) and dot-blot assays. From these results, we concluded that DAPF motif is the epitope of MAb D7. To our knowledge, this is the first report of a 4-mer epitope on the NS1 protein of AIV that can be recognized by MAb using a peptide microarray, which is able to simplify epitope identification, and that could serve as the basis for immune responses against avian influenza.

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