摘要
目的:从噬菌体展示肽库中,筛选可与肝癌细胞特异性结合的抗体模拟肽。方法:通过生物淘选使噬菌体富集。利用ELISA法,鉴定噬菌体单克隆原种的亲和性,并进行统计学分析。通过竞争ELISA,分析筛选所得抗体模拟肽的结合位点,并进一步分析抗体模拟肽的序列组成。结果:随着淘选次数的增加,出现噬菌体的富集。ELISA的结果显示,相对于正常肝细胞,筛选所得环状7肽对肝癌细胞系SMMC7721和BEL7402均有良好的结合活性(P<0.05),且与SMMC7721细胞的结合活性明显优于与BEL7402细胞的亲和性(P<0.05)。在α=0.01的水平上,7肽单克隆噬菌体原种可明显与scFv竞争结合SMMC7721细胞(0.005<P<0.01)。竞争性ELISA的结果显示,scFv在一定浓度(0～10-5)范围内,随着scFv浓度的逐渐降低,竞争抑制率也在逐渐减小。测序结果证实,所挑选的7肽阳性克隆的测序结果均相同。结论:通过淘选获得的抗体模拟肽(CLPRQSSFC)是有效的,为导向治疗提供了新的实验依据。
AIM: To screen the antibibody mimic peptide binding to hepatocellular carcinoma (HCC) from the circular 7-mer peptide libraries. METHODS: The enrichment of phage was achieved by repetitious biopanning. The affinity of peptide was evaluated by ELISA, and the binding site of the peptide was analysed by the competition ELISA . RESULTS: The output of phage gradually rose along with increasing of biopanning times. The obtained circular 7-mer peptide showed good binding activity to hepatocellular carcinoma cell lines SMMC-7721 and BEL-7402 (P<0.05), and its affinity to SMMC-7721 cells was superior to BEL-7402 (P<0.05) . At the level of α=0.01, the screened 7-mer peptide monoclonal stocks could obviously compete with scFv to bind SMMC-7721 cells (0.005<P<0.01). As the concentration of scFv gradually fell, the competition inhibition rate dropped accordingly. Sequence analysis showed that the sequences of 7-mer positive monoclonal stocks were identical. CONCLUSION: The peptide obtained through biopanning was specific and effective, which may provide further support for target therapy of HCC.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2005年第4期452-455,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助项目(No.30171091
30271478)
北京市自然科学基金资助(No.7022031)
关键词
噬菌体肽库
淘选
肝细胞癌
phage peptide library
biopanning
hepatocellular carcinoma
