期刊
JOURNAL OF INTEGRATIVE AGRICULTURE
卷 21, 期 7, 页码 2095-2105出版社
ELSEVIER SCI LTD
DOI: 10.1016/S2095-3119(21)63840-6
关键词
influenza A virus; nucleoprotein; monoclonal antibody; application
资金
- Natural Science Foun-dation of Heilongjiang Province, China [JQ2019C005]
- National Natural Science Foundation of China [31702265, 32172847]
In this study, two monoclonal antibodies (mAbs) against the nucleoprotein (NP) of influenza A virus (IAV) were generated and shown to have broad applications in virus research and disease diagnosis.
Monoclonal antibodies (mAbs) are widely used in virus research and disease diagnosis. The nucleoprotein (NP) of influenza A virus (IAV) plays important roles in multiple stages of the virus life cycle. Therefore, generating conserved mAbs against NP and characterizing their properties will provide useful tools for IAV research. In this study, two mAbs against the NP protein, 10E9 and 3F3, were generated with recombinant truncated NP proteins (NP-1 and NP-2) as immunogens. The heavy-chain subclass of both 10E9 and 3F3 was determined to be IgG2 alpha, and the light-chain type was kappa. Truncation and site-specific mutation analyses showed that the epitopes of mAbs 10E9 and 3F3 were located in the N terminal 84-89 amino acids and the C terminal 320-324 amino acids of the NP protein, respectively. We found that mAbs 10E9 and 3F3 reacted well with the NP protein of H1-H15 subtypes of IAV. Both 10E9 and 3F3 can be used in immunoprecipitation assay, and 10E9 was also successfully applied in confocal microscopy. Furthermore, we found that the 10E9-recognized (84)SAGKDP(89) epitope and 3F3-recognized (320)ENPAH(324) epitope were highly conserved in NP among all avian and human IAVs. Thus, the two mAbs we developed could be used as powerful tools in the development of diagnostic methods of IAV, and also surely promote the basic research in understanding the replication mechanisms of IAV.
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