山东医药2018，Vol.58Issue(24)：10-13,4.DOI:10.3969/j.issn.1002-266X.2018.24.003
噬菌体展示技术筛选胃癌高表达CD44分子特异性多肽序列的可行性分析
Feasibility of phage-display technology in screening specific polypeptide sequences in gastric cancer cells with high expression of CD44
摘要
Abstract
Objective To investigate the feasibility of phage-display technology in screening the specific polypeptide sequences in the gastric cancer cells with high expression of CD 44.Methods A phage-displayed random twelve-peptide library was used to screen CD44 purified protein for four rounds of affinity screening.Thirty clones were selected and were sequenced based on the single-strained DNA.ELISA was used to test the binding affinity of the selected phages to CD 44. The affinity of positive clones to CD44 integrated to the cell membrane was detected by immuno uorescence assay. Results During the four rounds, 30 clones were selected from the eluate of the 4th round of sequencing, and 7 sequences were determined.A six peptide motif which repeated 20 times was found in following analysis.Among the 7 sequences, 4 positive clones (S1, S2, S5 and S7) were identified by ELISA.The phage inserted sequence S1-WHXXXXXXQQA was identified, which had the highest affinity and specificity to CD 44.Conclusion Through phage-display technology, S1-WHXXXXXXQQA peptide sequence has the best affinity and specificity to CD 44, which may be a novel molecular probe for the molecular imaging diagnosis of gastric cancer.关键词
胃癌/噬菌体/噬菌体展示技术/多肽/CD44Key words
gastric carcinoma/phage/phage display technology/polypeptide/CD44分类
医药卫生引用本文复制引用
林小波,贾环,于照祥,李四光,李贞,常伟平..噬菌体展示技术筛选胃癌高表达CD44分子特异性多肽序列的可行性分析[J].山东医药,2018,58(24):10-13,4.基金项目
陕西省教育厅科学研究项目(17JK0660). （17JK0660）
