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Article

Identification of NP Protein-Specific B-Cell Epitopes for H9N2 Subtype of Avian Influenza Virus

by 1,2, 1,2, 1,2, 1,2, 1,2, 1,2 and 1,2,*
1
Key Laboratory of Animal Microbiology of China’s Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
2
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
*
Author to whom correspondence should be addressed.
Academic Editors: Chao-Nan Lin and Peck Toung Ooi
Viruses 2022, 14(6), 1172; https://doi.org/10.3390/v14061172
Received: 3 May 2022 / Revised: 25 May 2022 / Accepted: 26 May 2022 / Published: 28 May 2022
(This article belongs to the Topic Veterinary Infectious Diseases)
Avian Influenza (AI) caused by the H9N2 subtype of the avian influenza virus (AIV) poses a serious threat to both the poultry industry and to public health safety. NP is one of the major structural proteins in influenza viruses. B-cell determinants located on NP proteins have attracted increasing attention. In this study, based on the NP sequence of the H9N2 (A/chicken/Shandong/LY1/2017) strain, the truncated NP gene (71 AA–243 AA) was cloned and prokaryotically expressed in a pET-28a (+) vector. BALB/c mice were immunized with a purified recombinant of an NP protein to prepare a monoclonal antibody against NP proteins. The prokaryotic expression of four overlapping fragments, NP-N-96, NP-C-103, NP-C-54 and NP-C-49, were used to recognize an antigenic epitope of the NP protein. The results show that, after cell fusion, one hybridoma cell clone secreted the antibody specific to the NP protein, following screening with ELISA and indirect immunofluorescence, which is named the 4F5 monoclonal antibody (mAb). Western blotting on the overlapping fragments showed that the 230FQTAAQRA237 motif was identified as the minimal motif recognized by 4F5mAb, which was represented as the linear B-cell epitope of the NP protein. Homology analysis of this epitope shows that it was highly conserved in 18 AIVs analyzed in this study, and the epitope prediction results indicate that the epitope may be located on the surface of the NP protein. These results provide a strong experimental basis for studying the function of the NP protein of the H9N2 AIV and also strong technical support for the development of a universal assay based on an anti-NP monoclonal antibody. View Full-Text
Keywords: H9N2 avian influenza virus; NP protein; prokaryotic expression; monoclonal antibody; antigen epitope H9N2 avian influenza virus; NP protein; prokaryotic expression; monoclonal antibody; antigen epitope
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MDPI and ACS Style

Huang, X.; Huang, J.; Yin, G.; Cai, Y.; Chen, M.; Hu, J.; Feng, X. Identification of NP Protein-Specific B-Cell Epitopes for H9N2 Subtype of Avian Influenza Virus. Viruses 2022, 14, 1172. https://doi.org/10.3390/v14061172

AMA Style

Huang X, Huang J, Yin G, Cai Y, Chen M, Hu J, Feng X. Identification of NP Protein-Specific B-Cell Epitopes for H9N2 Subtype of Avian Influenza Virus. Viruses. 2022; 14(6):1172. https://doi.org/10.3390/v14061172

Chicago/Turabian Style

Huang, Xiangyu, Jingwen Huang, Guihu Yin, Yiqin Cai, Mengli Chen, Jianing Hu, and Xiuli Feng. 2022. "Identification of NP Protein-Specific B-Cell Epitopes for H9N2 Subtype of Avian Influenza Virus" Viruses 14, no. 6: 1172. https://doi.org/10.3390/v14061172

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