中国输血杂志2025，Vol.38Issue(8)：1023-1029,7.DOI:10.13303/j.cjbt.issn.1004-549x.2025.08.004
基于噬菌体展示技术鉴定与RhE抗原高亲和力的小分子多肽
Identification of small molecule peptides with high affinity for RhE antigen based on phage display technology
摘要
Abstract
Objective To identify small molecule peptides specifically binding to RhE antigen via phage display tech-nology,providing a theoretical basis to prevent RhE alloimmunization.Methods A peptide(FLWMFWPSVNSPLLR-SPIQRKNA)with RhE antigen-specific amino acids was artificially synthesized as the target peptide.A random phage dis-play 12-mer peptide library was screened in vitro against the artificially synthesized peptide via phage display technology.After two rounds screening in vitro and ELISA identification of the monoclonal phage,amplification and purification were carried out.Sequencing of the positive phage clones derived exogenous peptide amino acid sequences that specifically bind to RhE antigen,with target-binding specificity confirmed by competitive antibody inhibition assays.Results Two rounds screening of the target peptides results showed a significant enrichment of phages that specifically bind to the target peptides.96 phage clones were randomly selected for ELISA identification after two rounds screening,and the results showed 23 posi-tive clones with good affinity for the target peptide.After further verification by ELISA,13 phage clone results were consist-ent with the initial ELISA identification results,which indicated that these 13 clones have good affinity for the target pep-tide.DNA sequencing of these clones yielded five amino acid sequences:THDRNNTPVWRF,TPYETIFDPRTS,TFWQM-SADTQAL,AASSTLSIYPPR and SHDTRSPFTWGR.Competitive inhibition assays revealed all five peptides have competi-tive inhibitory effects with anti-E antibodies.Conclusion Based on phage display technology,5 small molecule peptides that specifically bind to RhE antigen have been successfully identified,which may provide a research basis for the preven-tion of HDFN by small molecule peptides and also provide new ideas for preventing RhE antigen alloimmunization.关键词
噬菌体展示技术/RhE抗原/红细胞同种免疫/小分子多肽/胎儿/新生儿溶血病Key words
phage display technology/RhE antigen/red blood cell(RBC)alloimmunization/small molecule peptides/haemolytic disease of the fetus and newborn(HDFN)分类
医药卫生引用本文复制引用
覃伟,李通,田力..基于噬菌体展示技术鉴定与RhE抗原高亲和力的小分子多肽[J].中国输血杂志,2025,38(8):1023-1029,7.基金项目
中国医学科学院医学与健康科技创新工程(2021-1-I2M-060) （2021-1-I2M-060）
