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为获得尤文肉瘤EWS-FLI1融合蛋白有效的HLA-A2.1限制性CTL表位,综合运用BIMAS,SYFPEITHI,Predep和IEDB方案对EWS-FLI1进行CTL表位的预测,得出8个CTL表位9肽序列;应用多项式方案、量化基序方案对8个表位9肽进一步进行筛选,获得其中4个9肽序列,并进一步运用分子模拟方法初步模拟了4个表位肽与HLA-A2.1分子间的相互结合作用;应用标准Fmoc方案合成CTL表位肽,RP-HPLC与MS分别鉴定合成肽的纯度与分子量;通过亲和力实验验证了表位肽QIQLWQFLL(EWS-FLI1304)与HLA-A2.1有较强的结合力,从而为表位肽的后继免疫学特性研究提供了基础.本研究提供了一个合理高效的表位筛选方法.
In order to obtain effective HLA-A2.1-restricted CTL epitopes of the Ewing’s sarcoma EWS-FLI1 fusion protein, the CTL epitopes of the EWS-FLI1 were predicted using the BIMAS, SYFPEITHI, Predep and IEDB protocols, and 8 CTL epitopes were obtained. 9 peptide sequences; using the polynomial scheme and quantification motif scheme to further screen the 8 peptides of 9 epitopes, obtained 9 peptide sequences, and further simulated the 4 epitope peptides and HLA-A2 using molecular simulation methods. The intermolecular interaction of 1 molecule was used to synthesize CTL epitope peptides using the standard Fmoc protocol. The purity and molecular weight of the synthesized peptides were identified by RP-HPLC and MS, respectively; the epitope peptides QIQLWQFLL (EWS-FLI1304) and HLA-A2 were verified by affinity experiments. .1 has a strong binding force, which provides the basis for subsequent immunological characteristics of epitope peptides. This study provides a rational and efficient method for epitope screening.