Screening specific minimum amino acid sequence triggering immunity to enterovirus 71
10.3760/cma.j.cn112309-20200107-00011
- VernacularTitle:筛选诱导肠道病毒71型特异性免疫应答的最短氨基酸基序
- Author:
Yan LIU
;
Wenchao GAO
;
Jialiang DU
;
Yueyue LIU
;
Qingchuan YU
;
Yan ZHAO
;
Rongrong ZHAO
;
Fei HAN
;
Xingliang FAN
;
Jiamei GAO
;
Tai GUO
- From:
Chinese Journal of Microbiology and Immunology
2021;41(1):45-49
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To screen the neutralizing epitope of enterovirus 71 (EV71) and determine the specific minimum amino acid sequence that triggers immunity for providing a theoretical basis for the development of synthetic peptide vaccines.Methods:EV71 neutralizing antibody-specific binding clones were panned and sequenced using a phage display random 12-peptide library to obtain the key sequences of neutralizing epitopes. A series of peptides containing the key sequences with N-terminal acetylation (AC) and C-terminal linking to Keyhole limpet hemocyanin (KLH) were synthesized. Serum samples were collected after immunizing mice with the modified peptides. Then the immunogenicity of the peptides and the neutralizing activity of serum samples were analyzed by Western blot, ELISA and neutralization test.Results:After three rounds of panning, cloning and sequencing, KQEKDL was identified as the key motif. The serum samples collected from the mice immunized with the modified series of peptides containing key motifs had different degrees of binding ability to EV71 and VP1 protein. The serum samples of mice immunized the synthetic peptide containing only the minimum key motif (AC-KQEKDL-KLH) had the strongest response to the other three peptides and EV71 and the highest neutralizing titer.Conclusions:The EV71 neutralizing epitope was successfully screened using the phage display random peptide library. The key motif of KQEKDL might be the specific minimum amino acid sequence that triggered the immune system. This study provides a theoretical basis for better understanding the immune response mechanism, evaluating the immunogenicity of the antigens and further research and development of polypeptide vaccines.
