中国免疫学杂志2011，Vol.27Issue(12)：1070-1074,5.DOI:10.3969/j.issn.1000-484X.2011.12.004
尤文肉瘤EWS-FLI1融合蛋白B淋巴细胞表位的预测及鉴定
Prediction and identification of B-lymphocyte epitopes derived from Ewing's sarcoma EWS-FLI1 fusion protein
摘要
Abstract
0bJ3ctive:To predict and identify 1he Ihym phocyle epitopes of Ew ing's saiconaEW S-FLIL fusion protein .M e1hods:The secondary structure and B-bin phocyte epitopes of EW S-FLH. Protein were can prehensivefy predicted by can prehensile algorithm ;epitope pep-tides w ere synihesized according to standard Fm oc proposal,purified by high perfbm ance liquid chran atography (H PLC ) and identified by m ass spectranetry(MS) to validate 1hemo]ecu]arweight.Antigenici1y of peptides and 1he sera titer of were evaluated by ELBA ;1he affinities of anti- peptides sera to EW S-FLH. Protein were identified by W estem bbt.ResulteiThreeB-iym phocyte epitopes were screened out according to can- prehensile algorithm .H PLC anafysis oonfhm ed 1he purity of any synthesized epitope peptides w as m ore 1han 85% at least and M S anafysis validated 1he m ofeukrw eights of 1hem accorded respective^ to 1heir theoretical values A111hree epitope peptides induced obvious antigen-antibody reactions,anongwhich epitope peptide2 (P2) did 1hemost;1heAbsorbance450 nm atlhe tineofP2 inmunoreactingwith rabbitantisenm after 3ri EW S-FLIL inmuniatfan increased to 1he apical value of 2 .46 atlhe reacting titer of 1=40 ,and 1he inmunoreactionwas still positive at 1he dilution titer of 1! 10 240 ; 1he titer of all 3 epitope peptides antisenm could increased gradual^ w ith 1he continuous increasing tin es of in- m uniation to N en Zealand white rabbit.The Absorbance 450 nm at 1he tin e of P2 inm unoreacting w ith rabbit antiserum after 4*1 P2 inm unia- tfanwas stilll .11 atlhe dilution titer of 1!512 000 .W estem blot analysis oonfhm ed 1hatanti-Pl and anti~P2 rabbit sera could bind to EW S- FLU protein .Conclusion :The Bhym phocyte epitopes peptides (PI and P2) derived Son EW S-FLH fusion protein of Ewing' s Sarcon a have potential antigenicity and inmunogenicily.关键词
尤文肉瘤/EWS-FLI1蛋白/融合蛋白/嵌合蛋白/B淋巴细胞表位Key words
Ew ing's sarcan a /EW S^LH protein /Fusion protein /Chin eric protein /Bhym phocyte epitopes分类
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黄路,韩智敏,罗嘉全,刘会文,张战民,廖翔,邓高荣,曹凯,安洪,舒勇..尤文肉瘤EWS-FLI1融合蛋白B淋巴细胞表位的预测及鉴定[J].中国免疫学杂志,2011,27(12):1070-1074,5.基金项目
本文为国家自然科学基金(30760254,81060221)、江西省自然科学基金(2009GQY0204)和江西省教育厅科研项目(GJJ08076)资助项目 （30760254,81060221）
