噬菌体随机肽库筛选MUC1抗原模拟表位  

Identification and screening of mimic epitope of MUC1

作　　者：杨清浩[1] 王祥卫[1] 金燕[1] 张立新[2] 

机构地区：[1]第三军医大学新桥医院泌尿外科,重庆400037 [2]北京军事医学科学院附属307医院肿瘤分子室,北京100850

出　　处：《第三军医大学学报》2005年第13期1332-1334,共3页Journal of Third Military Medical University

基　　金：国家"863"课题基金资助项目(2002AA214111)~~

摘　　要：目的噬菌体随机多肽文库筛选及鉴定MUC1抗原的模拟表位。方法利用纯化获得的BC2抗体筛选噬菌体随机7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30个阳性克隆,DNA序列分析并推导出氨基酸序列:TAPDLRP、SAPDLRP、AAPDSRP及LAPDFRP。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TAPDLRP、SAPDLRPAAPDSRP及LAPDFRP模拟MUC1抗原表位。Objective To identify and characterize the mimic epitope of MUC1 from random phage display peptide library. Methods The purified BC2 Ab was used to screen in phage random 7 peptide library. The positive clones were identified by sandwich ELISA and competitive inhibition assay. The homologous comparison of amino acid sequences of positive clones was conducted through searching the protein sequence databank on Internet. The digestion analysis of amino acid sequences was also conducted by a software. Results Thirty positive clones were acquired after 3 rounds of screening. Amino acid sequences deduced from DNA sequences showed four different sequences as TAPDLRP, SAPDLRP, AAPDSRP and LAPDFRP. 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 the control. Conclusion The results indicated that TAPDLRP, SAPDLRP, AAPDSRP and LAPDFRP are the mimotopes that can imitate the epitope of MUC1.

关 键 词：MCU1抗原 模拟表位 噬菌体随机7肽库 抗体 

分 类 号：R373.9[医药卫生—病原生物学] R392.11[医药卫生—基础医学]