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目的:从人肝癌T7 cDNA噬菌体展示肽库中筛选出能与人未成熟树突状细胞(iDC)特异结合的蛋白分子。方法:以人iDC为固相筛选目标,对人肝癌T7 cDNA噬菌体肽库进行4轮生物淘选,ELISA鉴定噬菌体单克隆。阳性克隆经PCR扩增并测序后行同源性比对分析。结果:4轮筛选富集现象不显著,获得80个只与人iDC特异性结合的阳性克隆。测序显示部分克隆序列相同,生物信息学比对发现与人iDC特异结合的阳性噬菌体克隆的同源蛋白中,可能与肝癌发生相关的分子有:人铁蛋白轻链、甲胎蛋白、α1微球蛋白前体、G蛋白偶联受体116和跨膜蛋白49等。结论:从人肝癌T7 cDNA噬菌体肽库中筛选获得了能与人iDC特异结合的分子,为阐明人肝癌发生发展过程中肿瘤免疫逃逸相关机制奠定了基础。
OBJECTIVE: To screen out the protein molecules that bind specifically to human immature dendritic cells (iDCs) from human hepatoma T7 cDNA phage display peptide library. METHODS: Human iDC was used as a solid phase screening target. Four rounds of biopanning of human hepatoma T7 cDNA phage peptide library were performed, and the phage monoclonal antibody was identified by ELISA. Positive clones were amplified by PCR and sequenced for homology analysis. Results: There was no significant enrichment in four rounds of screening, and 80 positive clones that specifically bound to human iDCs were obtained. Sequencing revealed some of the same cloned sequences. Among the homologous proteins found positive for phage clones that specifically bind to human iDCs, bioinformatics revealed that there may be related molecules of HCC: human ferritin light chain, α-fetoprotein, α1 microsphere Protein precursor, G protein coupled receptor 116 and transmembrane protein 49 and the like. Conclusion: The specific binding of human iDC to human Tc cDNA phage peptide library was obtained, which laid the foundation for elucidating the mechanism of tumor immune escape during the development of human hepatocellular carcinoma.