中国人兽共患病学报2024，Vol.40Issue(9)：807-813,7.DOI:10.3969/j.issn.1002-2694.2024.00.133
新型冠状病毒E蛋白B细胞表位的预测及鉴定
Prediction and identification of B-cell epitopes of SARS-CoV-2 E protein
摘要
Abstract
This work was aimed at predicting and verifying B-cell epitopes of SARS-CoV-2 E protein through bioinformatics methods,and clarifying the dominant B cell epitopes with mouse polyclonal antibody serum prepared through SARS-CoV-2 re-combinant E protein immunization and human positive serum vaccinated with inactivated SARS-CoV-2 vaccine.The structural and B-cell linear epitopes of SARS-CoV-2 E protein were predicted with SOPMA,Expasy,SWISS-MODEL,IEDB database,and Bepid-2.0 software.Candidate epitopes were expressed as GST-tagged recombinant protein fragments in an E.coli sys-tem,and their immunoreactivity with mouse and human poly-clonal positive serum against SARS-CoV-2 E protein was de-tected by western blotting and indirect ELISA,respectively.The epitope prediction results showed that E protein contained linear B cell epitopes Ser6-Val14 and Tyr57-Pro71,and the conformational epitopes of Glu8-Val12,Leu39-Tyr59,and Ser60-Leu65.The GST tagged recombinant E protein fragments of E1 and E3,containing Ser6-Val14 and Tyr57-Pro71 epitopes,respectively,as well as E2 without an epitope sequence as a control,were expressed in an E.coli expression system and successfully purified with an Ni-NTA column.Western blotting and indirect ELISA analysis indicated that all mouse and human SARS-CoV-2 positive sera positively reacted with only E1 and E3 proteins,but negatively reacted with E2 protein,thus indicating that the corresponding epitope prediction with Ser6-Val14 and Tyr57-Pro71 was correct.This study successfully predicted and preliminarily identified two linear B cell epitopes of SARS-CoV-2 E protein,thus providing a reference for the preparation of new coronavirus vaccines and the analysis of immune response characteristics.关键词
新型冠状病毒/B细胞表位/线性表位/预测/鉴定Key words
SARS-CoV-2/B-cell epitope/linear epitopes/prediction/identification分类
医药卫生引用本文复制引用
张鹏飞,刘钧,邹紫阳,康喜龙,宋丽,焦新安,孟闯,潘志明..新型冠状病毒E蛋白B细胞表位的预测及鉴定[J].中国人兽共患病学报,2024,40(9):807-813,7.基金项目
国家重点研发计划项目(No.2022YFC2604200)、江苏省重点研发计划项目(No.BE2021331)和江苏省第六期"333高层次人才培养工程"项目联合资助 Supported by the National Key Research and Development Program Project of(No.2022 YFC2604200),the Key Research and Development Program Project of Jiangsu Province(No.BE2021331),and the Sixth Phase of 333 High-level Talent Training Pro-ject of Jiangsu Province （No.2022YFC2604200）
