PDF(Pubmed)
Abstract：
African swine fever caused by African swine fever virus (ASFV) is an acute, highly contagious swine disease with high mortality. To facilitate effective vaccine development and find more serodiagnostic targets, fully exploring the ASFV antigenic proteins is urgently needed. In this study, the MGF_110-13L was identified as an immunodominant antigen among the seven transmembrane proteins. The main outer-membrane domain of MGF_110-13L was expressed and purified. Two monoclonal antibodies (mAbs; 8C3, and 10E4) against MGF_110-13L were generated. The epitopes of two mAbs were preliminary mapped with the peptide fusion proteins after probing with mAbs by enzyme-linked immunosorbent assay (ELISA) and Western blot. And the two target epitopes were fine-mapped using further truncated peptide fusion protein strategy. Finally, the core sequences of mAbs 8C3 and 10E4 were identified as 48WDCQDGICKNKITESRFIDS67, and 122GDHQQLSIKQ131, respectively. The peptides of epitopes were synthesized and probed with ASFV antibody positive pig sera by a dot blot assay, and the results showed that epitope 10E4 was an antigenic epitope. The epitope 10E4 peptide was further evaluated as a potential antigen for detecting ASFV antibodies. To our knowledge, this is the first report of antigenic epitope information on the antigenic MGF_110-13L protein of ASFV.
摘要：
非洲猪瘟由非洲猪瘟病毒(ASFV)引起,高死亡率的高传染性猪病。为了促进有效的疫苗开发并找到更多的血清诊断靶标，迫切需要充分探索ASFV抗原蛋白。在这项研究中,MGF_110-13L在7种跨膜蛋白中被鉴定为免疫显性抗原。表达并纯化了MGF_110-13L的主要外膜结构域。产生了针对MGF_110-13L的两种单克隆抗体(mAb;8C3和10E4)。通过酶联免疫吸附测定（ELISA）和Westernblot用mAb探测后，用肽融合蛋白初步定位了两个mAb的表位。并使用进一步截短的肽融合蛋白策略对两个目标表位进行精细定位。最后,单克隆抗体8C3和10E4的核心序列分别鉴定为48WDCQDGICKNKITESRFIDS67和122GDHQQLSIKQ131.合成表位肽，并用斑点印迹法检测ASFV抗体阳性猪血清，结果表明，表位10E4是一个抗原表位。进一步评估表位10E4肽作为检测ASFV抗体的潜在抗原。据我们所知,这是关于ASFV的抗原性MGF_110-13L蛋白的抗原性表位信息的首次报道。
