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Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex

机译:使用合成的肽表位-CpG-DNA-脂质体复合物针对靶向穗蛋白的MERS-CoV单克隆抗体的产生和表征

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摘要

Middle East respiratory syndrome coronavirus (MERS-CoV) uses the spike (S) glycoprotein to recognize and enter target cells. In this study, we selected two epitope peptide sequences within the receptor binding domain (RBD) of the MERS-CoV S protein. We used a complex consisting of the epitope peptide of the MERS-CoV S protein and CpG-DNA encapsulated in liposome complex to immunize mice, and produced the monoclonal antibodies 506-2G10G5 and 492-1G10E4E2. The western blotting data showed that both monoclonal antibodies detected the S protein and immunoprecipitated the native form of the S protein. Indirect immunofluorescence and confocal analysis suggested strong reactivity of the antibodies towards the S protein of MERS-CoV virus infected Vero cells. Furthermore, the 506-2G10G5 monoclonal antibody significantly reduced plaque formation in MERS-CoV infected Vero cells compared to normal mouse IgG and 492-1G10E4E2. Thus, we successfully produced a monoclonal antibody directed against the RBD domain of the S protein which could be used in the development of diagnostics and therapeutic applications in the future.
机译:中东呼吸综合征冠状病毒(MERS-CoV)使用刺突(S)糖蛋白识别并进入靶细胞。在这项研究中,我们选择了MERS-CoV S蛋白的受体结合域(RBD)内的两个表位肽序列。我们使用了由MERS-CoV S蛋白的表位肽和包裹在脂质体复合物中的CpG-DNA组成的复合物免疫小鼠,并产生了单克隆抗体506-2G10G5和492-1G10E4E2。蛋白质印迹数据表明,两种单克隆抗体均检测到S蛋白并免疫沉淀S蛋白的天然形式。间接免疫荧光和共聚焦分析表明该抗体对感染了MERS-CoV病毒的Vero细胞的S蛋白具有很强的反应性。此外,与正常小鼠IgG和492-1G10E4E2相比,506-2G10G5单克隆抗体显着减少了被MERS-CoV感染的Vero细胞中的噬斑形成。因此,我们成功生产了针对S蛋白RBD结构域的单克隆抗体,该抗体可用于未来的诊断和治疗应用开发。

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