利用单克隆抗体对非洲猪瘟病毒 pI215L 蛋白进行新型表位作图。
Novel Epitope Mapping of African Swine Fever Virus pI215L Protein Using Monoclonal Antibodies.
机构信息
Key Laboratory of Animal Diseases Diagnostic and Immunology, Ministry of Agriculture, MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
State Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou University, Lanzhou 730046, China.
出版信息
Viruses. 2023 Oct 12;15(10):2081. doi: 10.3390/v15102081.
The African swine fever virus (ASFV) is one of the most important pathogens that causes huge damage to worldwide swine production. The pI215L protein is found within the virion and expressed at a high level in infected porcine alveolar macrophages (PAMs), indicating a possible role of pI215L protein in ASFV detection and surveillance. In the present study, female BALB/c mice (5-6-week-old) were immunized with rpI215L protein, and six hybridomas, 1C1, 2F6, 2F10, 3C8, 5E1 and 5B3, steadily secreted anti-pI215L monoclonal antibodies (mAbs). Among them, 1C4, 5E1, and 5B3 had the IgG1 isotype with a Lambda light chain, 2F10 and 3C8 had the IgG1 isotype with a Kappa light chain, and 2F6 had the IgG2a isotype with a Kappa light chain. Western blot showed a good reactivity of the six mAbs against ASFV. Eight truncated polypeptides were produced for epitope mapping. Two novel B cell epitopes, 67LTFTSEMWHPNIYS80 and 167IEYFKNAASN176, were identified by the mAbs. Further analysis revealed that 2F6 mAb could be widely used in ASFV surveillance and 5B3 mAb might serve as a tool in the distinguishment of different ASFV genotypes. This study provides tools of monoclonal antibodies for further study of I215L function and contributes to the development of serological diagnosis and vaccine research.
非洲猪瘟病毒（ASFV）是一种重要的病原体，对全球养猪业造成了巨大的破坏。pI215L 蛋白存在于病毒粒子中，在感染的猪肺泡巨噬细胞（PAMs）中高水平表达，表明 pI215L 蛋白可能在 ASFV 的检测和监测中发挥作用。本研究中，雌性 BALB/c 小鼠（5-6 周龄）用 rpI215L 蛋白免疫，稳定分泌抗 pI215L 单克隆抗体（mAb）的 6 株杂交瘤细胞系 1C1、2F6、2F10、3C8、5E1 和 5B3。其中，1C4、5E1 和 5B3 为 IgG1 同种型，具有 Lambda 轻链，2F10 和 3C8 为 IgG1 同种型，具有 Kappa 轻链，2F6 为 IgG2a 同种型，具有 Kappa 轻链。Western blot 显示 6 株 mAb 对 ASFV 具有良好的反应性。通过 mAb 进行了表位作图，产生了 8 个截断多肽。鉴定了两个新的 B 细胞表位，67LTFTSEMWHPNIYS80 和 167IEYFKNAASN176。进一步分析表明，2F6 mAb 可广泛用于 ASFV 的监测，5B3 mAb 可能可用于区分不同的 ASFV 基因型。本研究为进一步研究 I215L 功能提供了单克隆抗体工具，并为血清学诊断和疫苗研究的发展做出了贡献。
Zotero 插件