人泌尿系上皮肿瘤疫苗的初步研究  

A study on epithelium tumor vaccine in urologic system

作　　者：王祥卫[1] 张立新[1] 张远宁[1] 叶钢[1] 张艮甫[1] 

机构地区：[1]第三军医大学附属新桥医院泌尿科,重庆400037

出　　处：《局解手术学杂志》2007年第5期302-304,共3页Journal of Regional Anatomy and Operative Surgery

基　　金：国家863课题资助项目(2002AA214111)

摘　　要：目的通过筛选噬菌体随机12肽库获得MUC1糖链抗原模拟表位。方法利用纯化获得的M a695抗体筛选噬菌体随机12肽库,通过夹心ELISA分析噬菌体克隆,测定阳性克隆DNA序列并进行同源性及氨基酸分析,竞争性抑制实验鉴定噬菌体克隆。结果经3轮筛选,获得了14个阳性克隆,DNA序列分析并推导出氨基酸序列:KHYDPFHHRMPQ,QADTARSVALAG,VPSKPDLHVRSI,MTPIHYWNHNRV。鉴定结果表明4个噬菌体展示肽克隆抑制率均在50%以上。结论所得序列KHYDPFH-HRMPQ,QADTARSVALAG,VPSKPDLHVRSI,及MTPIHYWNHNRV模拟了MUC1抗原表位。Objective To identify and characterize the mimic epitope of MUCI from phage display random 12 peptide library. Methods The purified Ma695 antibody was used to screen in phage display random 12 peptide library. The positive clones were identified by sandwich ELISA, competitive inhibition assay. The homologous comparison of amino acid sequences of positive clones was conducted through searching the protein sequence databank. The digestion analyses of amino acid sequences were also conducted by software. Results 14 positive clones were acquired after 3 rounds of screening. Amino acid sequences deduced from DNA sequences showed four different sequences as KHYDPFHHRMPQ, QADTARSVALAG, VPSKPDLHVRSI, MTPIHYWNHNRV. The inhibitory assay showed that the 4 mimic epitope peptides displaying on the phage surface could effectively inhibit the combination of antibody with antigen and the inhibitory rates of each mimic epitope were 50% higher than that in the control. Conclusion The results shows that KHYDPFHHRMPQ, QADTARSVALAG, VPSKPDLHVRSI, MTPIHYWNHNRV are the mimotopes which could mimic the epitope of MUCI.

关 键 词：MUC1抗原 模拟表位 噬菌体随机12肽库 

分 类 号：R730.3[医药卫生—肿瘤]