Generation and application of two monoclonal antibodies targeting conserved linear epitopes in the NP protein of influenza A virus

作     者：ZHAO Yu-hui WEN Xia LI Qi-bing JIANG Li WANG Guang-wen LIANG Li-bin WANG Xiu-rong CHEN Hua-lan LI Cheng-jun ZHAO Yu-hui;WEN Xia;LI Qi-bing;JIANG Li;WANG Guang-wen;LIANG Li-bin;WANG Xiu-rong;CHEN Hua-lan;LI Cheng-jun

作者机构：State Key Laboratory of Veterinary BiotechnologyHarbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069P.R.China 

出 版 物：《Journal of Integrative Agriculture》 (农业科学学报（英文版）)

年 卷 期：2022年第21卷第7期

页      面：2095-2105页

核心收录：

学科分类：090602[农学-预防兽医学] 09[农学] 0906[农学-兽医学] 

基　　金：supported by the Natural Science Foundation of Heilongjiang Province,China(JQ2019C005) the National Natural Science Foundation of China(31702265 and 32172847) 

主　　题：influenza A virus nucleoprotein monoclonal antibody application 

摘      要：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, 10 E9 and 3 F3, were generated with recombinant truncated NP proteins(NP-1 and NP-2) as immunogens. The heavy-chain subclass of both 10 E9 and 3 F3 was determined to be IgG2α, and the light-chain type was κ. Truncation and site-specific mutation analyses showed that the epitopes of mAbs 10 E9 and 3 F3 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 10 E9 and 3 F3 reacted well with the NP protein of H1–H15 subtypes of IAV. Both 10 E9 and 3 F3 can be used in immunoprecipitation assay, and 10 E9 was also successfully applied in confocal microscopy. Furthermore, we found that the 10 E9-recognized _(84) SAGKDP_(89) epitope and 3 F3-recognized 320 ENPAH324 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.