Abstract：
Akabane virus (AKAV) is characterized by abortion, stillbirth, premature birth, and congenital deformities in livestock and is widely distributed throughout Australia, Southeast Asia, East Asia, the Middle East, and Africa. Gc protein is the major neutralizing target of AKAV and is often considered as an immunogen to prepare neutralizing antibodies. In this study, we prepared and characterized three monoclonal antibodies (mAbs), 4D1, 4E6, and 4F12, against the Gc protein of AKAV (TJ2016 strain). Western blot (WB) and indirect immunofluorescence assay (IFA) analysis proved that the mAbs can react with both the truncated recombinant AKAV Gc protein and the natural Gc protein produced in the AKAV-infected cells. Further research demonstrated that these mAbs possess neutralizing activity. We next defined a neutralizing epitope 1134SVQSFDGKL1142 by screening a panel of overlapping peptides spanning the truncated Gc protein (aa991∼1232) using the generated neutralizing mAbs. Bioinformatic analysis shows that the neutralizing epitope is highly conserved across different genotypes of AKAV. The newly produced neutralizing mAbs and the identified neutralizing epitope in this study enrich the antigenic epitope information of the AKAV Gc protein and could have potential applications in the development of antigen and antibody detection systems that are specific to AKAV.
摘要：
Akabane病毒（AKAV）的特征是流产，死产,早产,和牲畜的先天性畸形，广泛分布在澳大利亚各地，东南亚,东亚,中东,和非洲。Gc蛋白是AKAV的主要中和靶标,并且通常被认为是制备中和抗体的免疫原。在这项研究中,我们制备并表征了三种单克隆抗体（mAb），4D1、4E6和4F12,针对AKAV(TJ2016株)的Gc蛋白。Westernblot（WB）和间接免疫荧光测定（IFA）分析证明，mAb可以与截短的重组AKAVGc蛋白和AKAV感染细胞中产生的天然Gc蛋白反应。进一步的研究证明这些mAb具有中和活性。接下来，我们通过使用产生的中和mAb筛选一组跨越截短的Gc蛋白（aa991~1232）的重叠肽来定义中和表位1134SVQSFDGKL1142。生物信息学分析显示中和表位在AKAV的不同基因型中是高度保守的。本研究中新产生的中和mAb和鉴定的中和表位丰富了AKAVGc蛋白的抗原表位信息，并且可能在开发对AKAV特异的抗原和抗体检测系统中具有潜在的应用。
