摘要
目的利用体内噬菌体展示技术筛选并鉴定与肺癌特异性结合的多肽。方法用肺癌细胞A549接种裸鼠复制荷瘤动物模型,将随机肽库尾静脉注入裸鼠体内,循环15 min后取肿瘤组织中结合的噬菌体,如此进行3轮体内筛选后挑取噬菌体克隆,ELISA初步鉴定噬菌体克隆对肺癌细胞的亲和力、特异性。将阳性噬菌体克隆扩增、测序获得外源多肽氨基酸序列,化学方法合成多肽,鉴定多肽对肺癌细胞和组织的亲和力、特异性。结果ELISA结果显示,随机挑选的20个噬菌体单克隆中,1个对A549具有很强亲和力。测序获得多肽,命名为zp2。化学合成多肽zp2。竞争抑制、细胞免疫荧光和组织免疫荧光实验结果表明多肽zp2与肺癌细胞A549及肺癌组织特异性结合。结论利用体内噬菌体展示技术筛选出与肺癌细胞A549特异性结合的多肽,为肺癌诊断及治疗药物的研制奠定基础。
To obtain the polypeptides specifically binding to lung cancer from peptide libraries by using in vivo phage display technology.Nude mice were inoculated with human lung tumor cells A549 for setting up tumor-bearing nude mouse model.The Ph.D.-12 Phage Display Peptide Library was injected intravenously via tail vein,and 15 min after the injection,the mice were sacrificed and the phage was rescued from tumor tissues.The phage recovered from the tissues was amplified and purified,and then re-injected for next round screening.After 3 rounds of in vivo screening,the phage peptides homed to the tumor tissues was obtained.Then the phage clones were preliminary identified by cell enzyme-linked immunosorbent assay（ELISA） on lung cancer cells affinity and specifcity.The positive phage clone was amplified and sequenced.Then the peptide was synthesized by chemical methods and identified for the affinity and specificity to lung cancer cells and tissues.The 20 phage clones were identified by ELISA,one of them was specially bound to the A549 cell line;the amino acid sequence was deduced and the peptide was synthesized,and named zp2,which were specially bound to the A549 cell line and the lung cancer tissue.All the results lay the foundation for the development of diagnostic reagents and drugs of lung cancer.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2010年第1期68-71,共4页
Immunological Journal
基金
教育部重点基金资助(208105)
广东药学院人才启动基金(2006)
广东药学院师资队伍建设经费"中青年骨干教师"专项经费资助
