Identification of Two Novel Linear Neutralizing Epitopes within the Hexon Protein of Canine Adenovirus Using Monoclonal Antibodies

文献类型： 外文期刊

第一作者： Wang, Shujie

作者： Wang, Shujie;Ren, Xiao;Xue, Wenjiao;He, Haijuan;Wang, Hongfeng;Wang, Gang;Cai, Xuehui;Wang, Chunsheng;He, Haijuan;Zhu, Yanzhu

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关键词： canine adenovirus; identification; hexon protein; monoclonal antibodies; novel epitopes

期刊名称：VACCINES （ 影响因子：4.422； 五年影响因子：5.513 ）

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年卷期： 2021 年 9 卷 2 期

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收录情况： SCI

摘要： Canine adenovirus (CAdV) has a high prevalence in canine populations. High affinity neutralizing antibodies against conserved epitopes can provide protective immunity against CAdV and protect against future outbreaks. In this study, we identified two CAdV-2-specific neutralizing monoclonal antibodies (mAbs), 2C1 and 7D7, which recognized two linear-dependent epitopes. MAb 2C1 potently neutralized CAdV-2 with a 50% neutralization titer (NT50) of 4096, and mAb 7D7 partially neutralized CAdV-2 with a 50% NT50 of 64. Immunoprecipitation, Western blot and protein spectral analysis indicated that both neutralizing mAbs recognized the hexon protein (Hex) of CAdV-2. Through a 12-mer random peptide phage display and synthetic peptides analysis, we finely mapped the neutralizing epitopes to two 10-amino acid (aa) peptides within the CAdV Hex: (634)RIKQRETPAL(643) located on the surface region; and (PESYKDRMYS745)-P-736 located in the inner region of the expected 3D structure of trimeric Hex. Importantly, the two epitopes are highly conserved among all CAdV isolates by sequence alignment analysis. Thus, these results provide insights into the interaction between virus and mAbs at the aa level and may have potential applications in the development of novel therapeutic or epitope-based vaccines, antibody therapeutics and a diagnostic method suitable for the rapid detection of all CAdVs.

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