识别9E10单抗的噬菌体短肽的免疫学特性分析

STUDIES ON THE IMMUNOLOGICAL PROPERTIES OF PHAGE PEPTIDES RECOGNIZED BY McAb 9E10

为分析从单价表达短肽库中筛选到的两种克隆的免疫学特性，将单价噬菌体短肽表达载体中的ｇⅢ片段切除，在大肠杆菌中表达可溶性短肽分子，ＥＬＩＳＡ竞争抑制实验证实，可溶性短肽分子可竞争抑制９Ｅ１０单抗与Ｃ－ｍｙｃ＋肽结合。进一步将噬菌体短肽的单价表达载体改建为多价表达载体，分别免疫兔和小鼠，制备免疫抗血清，ＥＬＩＳＡ实验和竞争抑制实验结果证实，两种克隆的免疫抗血清均可与９Ｅ１０单抗所识别的抗原表位结合，表明这两个克隆尽管在序列上与原自然抗原不同，但其在结构上真正模拟了ｃ－ｍｙｃ＋肽的抗原表位，在体内可诱发相似的免疫反应。

The immunological properties of two phage peptide clones isolated by screening a phage peptide library against McAb 9E10 were analyzed.Soluble peptide was prepared by removing gene Ⅲ fragment from the expression vector.The induced supernatant exhibited capacity to bind 9E10 demonstrating the short peptide could be secreted and retained its biological activity.The immunogenicity of the phage peptides were further investigated by construction of multivalency phage peptides and immunization of animals with the phage peptides.ELISA assay and competitive ELISA showed that antiserum from both mice and rabbit immunized with either clone could bind to the original antigen,cmyc decapeptide.These results denoted that in spite of dissimilarity of the peptides with cmyc decapeptide,they are capable of inducing similar immunoresponse in vivo,thus actually mimicked the antigenic epitope.