Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon 24252, Korea.
Department of Microbiology, College of Medicine, Hallym University, Chuncheon 24252, Korea.
BMB Rep. 2019 Jun;52(6):397-402. doi: 10.5483/BMBRep.2019.52.6.185.
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. [BMB Reports 2019; 52(6): 397-402].
中东呼吸综合征冠状病毒(MERS-CoV)利用刺突(S)糖蛋白识别并进入靶细胞。在本研究中,我们选择了 MERS-CoV S 蛋白受体结合域(RBD)内的两个表位肽序列。我们使用包含 MERS-CoV S 蛋白表位肽和包封在脂质体复合物中的 CpG-DNA 的复合物免疫小鼠,产生了单克隆抗体 506-2G10G5 和 492-1G10E4E2。Western blot 数据表明,两种单克隆抗体均检测到 S 蛋白,并免疫沉淀了 S 蛋白的天然形式。间接免疫荧光和共聚焦分析表明,这些抗体对感染 MERS-CoV 病毒的 Vero 细胞中的 S 蛋白具有强烈的反应性。此外,与正常小鼠 IgG 相比,506-2G10G5 单克隆抗体显著降低了感染 MERS-CoV 的 Vero 细胞中的斑块形成。因此,我们成功地产生了针对 S 蛋白 RBD 域的单克隆抗体,可用于未来诊断和治疗应用的开发。[BMB 报告 2019;52(6):397-402]。