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目的利用噬菌体展示肽库筛选出与肺癌NCI-H1299细胞特异性结合的多肽,鉴定其特异性和亲和力,为寻找肺癌早期诊断和治疗标志物打下基础。方法以肺癌细胞NCI-H1299为靶细胞,肺二倍体成纤维细胞MRC-5为吸附细胞,在37℃下对噬菌体随机十二肽库进行三轮减性筛选,挑取单克隆ELISA初步鉴定噬菌体克隆亲和力;阳性克隆提取质粒,测序后合成多肽,鉴定多肽的亲和力及特异性。结果经ELISA初步鉴定,随机挑选的20个单克隆中,其中6号对NCI-H1299亲和力最高,将其命名为PhageZS6。细胞免疫荧光试验进一步证明,合成的相应多肽FITCZS6对NCI-H1299具有高亲和力。结论利用噬菌体随机十二肽成功筛选出与肺癌细胞具有较高亲和力的多肽ZS6,为肺癌的早期诊断和靶向治疗奠定基础。
OBJECTIVE: To screen out the specific binding peptides of NCI-H1299 cells from phage display peptide library and identify its specificity and affinity, so as to lay the foundation for the early diagnosis and treatment of lung cancer. Methods Lung cancer cell NCI-H1299 was used as target cells and lung diploid fibroblasts MRC-5 was used as adherent cells. The phage randomized 12-mer peptide library was screened by three rounds at 37 ℃ for the first time. Phage cloning affinity; positive clones plasmid extraction, synthesis of peptides after sequencing to identify the affinity and specificity of the polypeptide. The results of the initial identification by ELISA, randomly selected 20 monoclonal, 6 of which NCI-H1299 highest affinity, named PhageZS6. Cellular immunofluorescence assay further demonstrated that the synthesized corresponding polypeptide FITCZS6 has high affinity to NCI-H1299. Conclusion The peptide ZS6 with high affinity to lung cancer cells was successfully screened by phage randomized dodecyl peptide, which laid the foundation for the early diagnosis and targeted therapy of lung cancer.