通过噬菌体筛选表达人CD59的CHO细胞结合的内化短肽

SCREENING THE SHORT INTERNATION PEPTIDE COMBINED WITH CHO CD59 THROUGH PHAGE

目的筛选并鉴定与表达人CD59的中国仓鼠卵巢(CHO)细胞特异结合的内化短肽,为设计具有拮抗CD59肿瘤逃逸活性的短肽药物奠定基础。方法以高表达人CD59的CHO细胞为靶,对噬菌体随机十二肽库进行5轮亲和筛选,并进行竞争结合实验,用ELISA法筛选噬菌体阳性克隆并对其进行DNA序列分析。结果随机挑选的10个克隆中有6个与人CD59结合力高,测序后得到2个高度同源的氨基酸序列。结论获得了与高表达人CD59的CHO细胞结合的内化短肽序列,为进一步设计与肿瘤逃逸相关的CD59活性位点的短肽药物提供了参考依据。

Objective To search for a short internation peptide specifically combined with CHO cells for the purpose to establish a base of designing a peptide drug to inhibit CD59 on tumor sneaking through. Methods Taking CHO cells of high expressing CD59 as a target, a five-round affinity screening was done randomly for peptide library 12, After competitive test, positive phage clones were selected by ELISA and subsequently underwent DNA sequencing. Results Six out of 10 selected phage clones possessed the high affinity to CD59. After DNA sequencing, two pieces of short peptides highly homogeneous to CD59 were obtained. Conclusion This study might provide the reference for the design of a drug similar to the short peptide of CD59 on tumor sneaking through by obtaining the sequence of the internation peptide.