噬菌体随机肽库淘选肺癌细胞NCI-H1299特异性结合多肽

Panning and identification of lung cancer cell line NCI-H1299 specific peptide from a phage display peptide library

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。

Aim To obtain polypeptides which could specifically bind to NCI-H1299 cell line from peptide libraries, and to further identify and verify these polypeptides for the screening and identifying of the novel lung cancer markers in early diagnosis and treatment of lung cancer. Methods The lung cancer NCI-H1299 cell line was used as the antigen and MRC-5 was used as control for subtraction biopanning from a phage display peptide library at 37℃. The positive and specific binding clones were identified by cell-based ELISA and immunocytochemical staining, and the identified clones was sequenced. Thus the amino acid sequence was deduced and the peptide was synthesized. The peptide was identified by immunofluorescence. Results Through a cell-based ELISA, immunocytochemical staining, and immunofluorescence, the Phage ZS 6 and synthetic peptide ZS 6 were shown to specially bind to the NCI-H1299. Conclusion A peptide specific binding to lung cancer cell line NCI-H1299 has been selected from phage display peptide libraries. Therefore, it provides a potential tool for early diagnosis of lungcancer or targeted drug delivery in chemotherapy.