Preparation and immunogenicity evaluation of two PreF trimer recombinant protein vaccines against respiratory syncytial virus
10.3760/cma.j.cn112309-20240112-00024
- VernacularTitle:两种呼吸道合胞病毒PreF三聚体重组蛋白疫苗的制备和免疫原性评价
- Author:
Heng ZHANG
1
;
Hongjian XIAO
;
Haiwei LI
;
Yaoyun YANG
;
Jinmei DUAN
;
Zhihua LI
Author Information
1. 中国医学科学院医学生物学研究所，病原体感染防控教育部重点实验室（北京协和医学院），昆明 650031
- Keywords:
Respiratory syncytial virus;
PreF trimer protein;
Antibody titer
- From:
Chinese Journal of Microbiology and Immunology
2024;44(2):101-109
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct and purify four respiratory syncytial virus (RSV) PreF proteins through gene sequence design and optimization and evaluate their immunogenicity.Methods:Coronin-1A and T4 trimer protein gene sequences were optimized with Human and CHO codons, and then added to RSV F protein sequence. The above plasmids were transfected into Expi293F cells for protein expression. After purification by nickel column, four trimer proteins were prepared. SDS-PAGE and Western blot were performed for protein identification. BALB/c mice were immunized at week 0 and week 3, and blood samples were collected to measure the activities of binding and neutralizing antibodies in serum.Results:SDS-PAGE and Western blot showed that the four proteins had stable trimer structure. Antigen-antibody affinity test showed that the four trimer proteins had strong affinity with RSV-specific monoclonal antibodies 8897, D25, Motavizumab, AM14 and Palivizumab. The titers of antibodies induced by the two T4 trimers were higher after the initial immunization, while there was a substantial increase in the titers of antibodies induced by Human codon-optimized trimer protein after the second immunization.Conclusions:PreF trimer protein can be prepared by adding any of the two different heterotrimer motifs, and induce effective binding and neutralizing antibodies in mice.
