南京医科大学学报（自然科学版）2013，Vol.33Issue(5)：593-597,5.DOI:10.7655/NYDXBNS20130506
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
Preparation and immunogenicity characteristics analysis of a epitope fusion protein of Epstein-Bar virus encoded latent membrane protein 2A
摘要
Abstract
Objective:To prepare a immunogenic epitope tandem of latent membrane protein 2A (LMP2A) of Epstein-Ban: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.关键词
鼻咽癌/EB病毒/潜伏膜蛋白2A/多克隆抗体/重组基因Key words
nasopharyngeal carcinoma/ Epstein-Barr virus/ latent membrane protein 2A/polyclonal antibody/ recombinant gene分类
医药卫生引用本文复制引用
曹清,唐小军,李文杰,熊四平,毛园,熊林,刘玉,王长军,冯振卿..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）,2013,33(5):593-597,5.基金项目
南京医科大学科技发展基金重点项目(NY2005 D2D13) （NY2005 D2D13）
江苏省自然科学基金(BK2008481) （BK2008481）
江苏省科技厅社会发展支撑计划项目(BE2009152) （BE2009152）
