Identification of small molecule peptides with high affinity for RhE antigen based on phage display technology
10.13303/j.cjbt.issn.1004-549x.2025.08.004
- VernacularTitle: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
- Author:
Wei QIN
1
;
Tong LI
1
;
Li TIAN
2
Author Information
1. Pengzhou Second People's Hospital, Pengzhou 611900, China
2. Key Laboratory of Transfusion Adverse Reactions, Chinese Academy of Medical Sciences, Institute of Blood Transfusion, Chinese Academy of Medical Sciences&Peking Union Medical College, Chengdu 610052, China
- Publication Type:Journal Article
- Keywords:
phage display technology;
RhE antigen;
red blood cell (RBC) alloimmunization;
small molecule peptides;
haemolytic disease of the fetus and newborn (HDFN)
- From:
Chinese Journal of Blood Transfusion
2025;38(8):1023-1029
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To identify small molecule peptides specifically binding to RhE antigen via phage display technology, providing a theoretical basis to prevent RhE alloimmunization. Methods: A peptide (FLWMFWPSVNSPLLRSPIQRKNA) with RhE antigen-specific amino acids was artificially synthesized as the target peptide. A random phage display 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 positive clones with good affinity for the target peptide. After further verification by ELISA, 13 phage clone results were consistent with the initial ELISA identification results, which indicated that these 13 clones have good affinity for the target peptide. DNA sequencing of these clones yielded five amino acid sequences: THDRNNTPVWRF, TPYETIFDPRTS, TFWQMSADTQAL, AASSTLSIYPPR and SHDTRSPFTWGR. Competitive inhibition assays revealed all five peptides have competitive 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 prevention of HDFN by small molecule peptides and also provide new ideas for preventing RhE antigen alloimmunization.
