摘要
目的:利用噬菌体7肽库进行体外快速差减筛选(biopanning and rapid analysis of selective interactive ligands,BRASIL),从而获得卵巢癌细胞株HO-8910细胞表面特异性结合肽。方法:以中国仓鼠卵巢细胞株CHO为吸附细胞,卵巢癌细胞株HO-8910为靶细胞,利用噬菌体展示技术联合差减筛选策略,经过连续5轮生物淘洗,随机挑选13个噬菌体单克隆进行DNA测序,利用噬菌体竞争结合实验、ELISA实验、细胞免疫染色、细胞免疫荧光、合成多肽的竞争抑制实验鉴定阳性噬菌体的亲和性及特异性。结果:筛选出的噬菌体NPMIRRQ与卵巢癌细胞株HO-8910有较高的亲和性及特异性。结论:阳性噬菌体表面展示的多肽NPMIRRQ可能为卵巢癌的早期诊断和转移复发监测提供新思路。
（Objectives：To screen the specific peptide binding on the surface of ovarian cell line HO-8910 by phage peptide library using biopanning and rapid analysis of selective interactive ligands（ BRASIL） method.Methods：Utilizing the BRASIL method to screen the 7-mer phage display peptide library on two cell lines,chinese hamster ovarian cell line CHO and human ovarian tumor cell line HO-8910.After five rounds of in vitro re-iterative panning,thirteen blue plaques from a tittering plate of the fifth round were randomly selected for sequencing of DNA.Phage competitive experiment,ELISA,immunocytochemistry studies,immunofluorescence studies and competitive inhibition assay were applied to confirm the affinity and specificity of the positive phage clone binding to HO-8910 cells.Results：The screened phage clone NPMIRRQ showed higher affinity and specificity combining with HO-8910 cells.Conclusions：The peptide NPMIRRQ displayed on the surface of positive phage perhaps provide a new direction for early diagnosis,metastasis monitoring and treatment of ovarian tumor.
出处
《中国性科学》
2016年第1期56-60,共5页
Chinese Journal of Human Sexuality
基金
国家自然科学基金项目(81172463)
浙江省温州市科技局资助项目(Y20140319)
关键词
卵巢癌
噬菌体展示技术
噬菌体多肽库
体外快速差减筛选技术
多肽
Ovarian cancer
Phage display
Phage peptide library
Biopanning and rapid analysis of selec tive interactive ligands（BRASIL）
Peptides
