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目的以肺癌血清从噬菌体展示肽库中筛选出能够用于血清学筛查和肺癌病理分型的多肽。方法利用M13噬菌体随机12肽展示肽库对24例临床确诊肺癌患者的血清和24例体检者的血清进行4轮吸附-洗脱的筛选。挑取筛选后的噬菌体单克隆,皆以血清和细胞株进行试验,选出特异性及确诊率较高的单克隆。目标单克隆通过DNA测序确定其所表达的多肽序列,进而人工合成带FITC标记的多肽,以肺癌细胞、正常肺细胞和肺癌组织切片进行荧光成像检测。结果随机挑取30个经过4轮筛选的噬菌体单克隆,通过血清和细胞株的试验,筛选出6个特异性和确诊率较高的单克隆,其中单克隆ZZ-8肺癌血清确诊率达到41.6%,且鳞癌的比例较高。ZZ-8经DNA测序分析,确定其表达的多肽序列为:APTPSTQMNLLP。FITC标记的多肽在细胞和组织切片的荧光成像检测中,均显示良好的特异性。结论人工合成的多肽ZZ-8能够为研发肺癌诊断试剂奠定基础。
Objective To screen the peptides from phage display peptide library of lung cancer serum for serological screening and pathological classification of lung cancer. Methods Serum samples of 24 patients with clinically diagnosed lung cancer and 24 sera from 24 random samples of M13 phage display peptide library were screened for 4 rounds of adsorption and elution. Pick the selected phage monoclonal, all with serum and cell lines were tested to select a specific and high-confirmed monoclonal. The target monoclonal DNA was sequenced to determine the sequence of the expressed polypeptide. Then the FITC-labeled polypeptide was synthesized and detected by fluorescence imaging with lung cancer cells, normal lung cells and lung cancer tissue sections. Results Thirty randomly selected phage clones were screened by four rounds. Six of them were screened out by the experiments of serum and cell lines. The results showed that six monoclonal antibodies with high specificity were detected, of which the diagnosis rate of monoclonal ZZ-8 lung cancer was 41.6 %, And a higher proportion of squamous cell carcinoma. ZZ-8 DNA sequencing analysis to determine the expression of the polypeptide sequence: APTPSTQMNLLP. FITC-labeled polypeptides showed good specificity in fluorescence imaging of cell and tissue sections. Conclusion The synthetic peptide ZZ-8 can lay the foundation for the development of diagnostic reagents for lung cancer.