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利用单克隆抗体鉴定非洲猪瘟病毒MGF_110-13L蛋白上的两个线性表位

Identification of Two Linear Epitopes on MGF_110-13L Protein of African Swine Fever Virus with Monoclonal Antibodies.

作者信息

Zhang Shu-Jian, Niu Bei, Liu Shi-Meng, Zhu Yuan-Mao, Zhao Dong-Ming, Bu Zhi-Gao, Hua Rong-Hong

机构信息

State Key Laboratory of Animal Disease Control and Prevention, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150069, China.

Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.

出版信息

Animals (Basel). 2024 Jul 1;14(13):1951. doi: 10.3390/ani14131951.

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 WDCQDGICKNKITESRFIDS, and GDHQQLSIKQ, 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

由非洲猪瘟病毒(ASFV)引起的非洲猪瘟是一种急性、高传染性且死亡率高的猪病。为推动有效的疫苗研发并找到更多血清学诊断靶点,迫切需要全面探索ASFV抗原蛋白。在本研究中,MGF_110-13L被鉴定为七个跨膜蛋白中的免疫显性抗原。表达并纯化了MGF_110-13L的主要外膜结构域。制备了两种针对MGF_110-13L的单克隆抗体(mAb;8C3和10E4)。通过酶联免疫吸附测定(ELISA)和蛋白质印迹法用单克隆抗体探测后,用肽融合蛋白初步绘制了两种单克隆抗体的表位。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了精细绘制。最后,单克隆抗体8C3和10E4的核心序列分别被鉴定为WDCQDGICKNKITESRFIDS和GDHQQLSIKQ。合成了表位肽并用ASFV抗体阳性猪血清通过斑点印迹法进行探测,结果表明表位10E4是一个抗原表位。进一步评估了表位10E4肽作为检测ASFV抗体的潜在抗原。据我们所知,这是关于ASFV抗原性MGF_110-13L蛋白的抗原表位信息的首次报道。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09db/11240426/2019a3007298/animals-14-01951-g001.jpg

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