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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SKOV3细胞表面特异性结合短肽。方法:以中国仓鼠卵巢细胞株CHO为吸附细胞,卵巢癌细胞株SKOV3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100个噬菌体单克隆进行ELISA检测,选取读数>0.6的单克隆进行DNA测序并序列分析,选择并合成相应短肽,经细胞免疫荧光验证短肽和卵巢癌细胞SKOV3结合的亲和性及特异性。结果:筛选出的短肽AFSGSLP与卵巢癌细胞株SKOV3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AFSGSLP可能为卵巢癌的早期诊断和转移复发监测提供新思路。
OBJECTIVE: To obtain the surface-specifically bound short peptides of ovarian cancer cell line SKOV3 by using the phage 7 peptide library for biopanning and rapid analysis of selective interactive ligands (BRASIL). Methods: Using Chinese hamster ovary cell line CHO as the adherent cells and ovarian cancer cell line SKOV3 as the target cells, phage display technology combined with the subtractive screening strategy was used to select 100 phage clones for ELISA , Selecting a single clone with the reading> 0.6 for DNA sequencing and sequence analysis, selecting and synthesizing the corresponding short peptide, and verifying the affinity and specificity of the short peptide and the ovarian cancer cell SKOV3 by immunofluorescence. RESULTS: The selected AFSGSLP peptide had high affinity and specificity with ovarian cancer cell line SKOV3. Conclusion: AFSGSLP displayed on the surface of positive phage may provide new ideas for the early diagnosis and metastasis of ovarian cancer.