Identification of catalytically active domain epitopes in neuraminidase protein of H9N2 subtype of avian influenza virus

被引:0
|
作者
Huang, Xiangyu [1 ,2 ]
Cai, Yiqin [1 ,2 ]
Yin, Guihu [1 ,2 ]
Chen, Zili [3 ]
Hu, Jianing [1 ,2 ]
Gao, Zichen [1 ,2 ]
Guo, Xinyu [1 ,2 ]
Xiong, Fuqiang [1 ,2 ]
Feng, Xiuli [1 ,2 ,4 ]
机构
[1] Nanjing Agr Univ, Coll Vet Med, Chinas Minist Agr, Key Lab Anim Microbiol, Nanjing, Peoples R China
[2] Nanjing Agr Univ, Coll Vet Med, MOE Joint Int Res Lab Anim Hlth & Food Safety, Nanjing, Peoples R China
[3] Rudong Agr & Rural Affairs Bur, Agr Comprehens Law Enforcement Brigade Rudong, Rudong, Peoples R China
[4] Nanjing Agr Univ, Coll Vet Med, Chinas Minist Agr, Key Lab Anim Microbiol, Nanjing 210095, Peoples R China
基金
中国国家自然科学基金;
关键词
NA protein; monoclonal antibody; antigen epitope; catalytically active domain; epitope scanning technique; TRANSMEMBRANE DOMAIN; A VIRUS; GENESIS;
D O I
10.1080/03079457.2023.2239191
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
H9N2 subtype of avian influenza virus (AIV) is primarily a bird virus, which is widespread in clinical avian disease, and reported in cases of human infection. As one of the surface proteins of AIV, the neuraminidase (NA) protein plays an important role mainly in viral budding. However, vaccine development and detection methods for NA of H9N2 AIVs are in urgent clinical need. In this study, a truncated NA gene (205-900 bp) was cloned from the NA sequence of H9N2 strain, and then expressed using pET-28a (+) vector. This purified recombinant NA protein was used to immunize BALB/c mice, and the monoclonal antibodies were screened through the indirect enzyme-linked immunosorbent assay (ELISA). Next, eight prokaryotic expression vectors were constructed for epitope identification. After cell fusion, three hybridoma cell lines producing the antibodies special to NA protein were screened by ELISA, western blotting, and indirect immunofluorescence; these were named 1B10, 2B6, and 5B2, respectively. Epitope scanning techniques were used to identify three B-cell epitopes recognized by these three monoclonal antibodies, (196)KNATASIIYDGMLVD(210), (210)DSIGSWSKNIL(220) and (221)RTQESECVCI(230). The subsequent homology analysis revealed the three epitopes were highly conserved in H9N2 AIV strains. The structural predictions of the antigenic epitopes indicated that all three epitopes were located in the catalytic region of NA. These results provide a basis for studying the function of the NA protein of H9N2 AIV and technical support for the development of a universal detection method based on anti-NA monoclonal antibodies.
引用
收藏
页码:377 / 387
页数:11
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