Preparation and immunogenicity characteristics analysis of a epitope fusion protein of Epstein-Bar virus encoded latent membrane protein 2A
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    Abstract:

    Objective:To prepare a immunogenic epitope tandem of latent membrane protein 2A (LMP2A) of Epstein-Barr virus and analyze its characteristics of immunology. Methods:LMP2A epitopes were analyzed by using DNAstar software,and two immunogenic epitopes were selected due to stronger immunogenicity. The genes of these two epitopes were integrated through gene synthesis,and then cloned into prokaryotic expression vector pET28a. The recombinant epitopes were expressed and purified by E.coli BL21,and then the protein was utilized to immunize mouse to prepare anti-epitope fusion protein polyclonal antibody after the analysis of SDS-PAGE and Western blot assay. The titer of the antibody was assessed by ELISA and the immunohistochemical experiment was performed to test the specificity of the polyclonal antibody for natural LAMP2A. Results:Through prokaryotic expression and purification,high purity fusion protein was obtained. By mouse immunization,anti-LMP2A polyclonal antibody of high titer and specificity was prepared,which can be applied to ELISA and immunohistochemical analysis. Conclusion:An epitope fusion protein,which shares the natural antigen immunogenicity,can be used to prepare the polyclonal antibody specific for LMP2A. This study has laid a solid foundation for screening whole humanized genetic engineering antibody by epitope fusion protein.

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曹 清,唐小军,李文杰,熊四平,毛 园,熊 林,刘 玉,王长军,冯振卿,陈仁杰. EBV编码潜伏膜蛋白2A抗原表位的串联表达及其免疫原性分析[J].南京医科大学学报(自然科学版英文版),2013,(5):593-597.

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  • Received:January 06,2013
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  • Online: May 29,2013
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