■中东呼吸综合征冠状病毒是一种高致病性病毒,对公众健康构成重大威胁。
■本研究的目的是开发和表征靶向中东呼吸综合征电晕病毒(MERS-CoV)的刺突蛋白S1亚基的新型小鼠单克隆抗体。
■在这项研究中,使用杂交瘤技术产生并表征三种针对MERS-CoV的小鼠单克隆抗体(mAb)。评价mAb的反应性和中和活性。通过杂交瘤技术通过融合来自MERS-CoV-S1免疫小鼠的骨髓瘤细胞和脾细胞来产生mAb。使用酶联免疫吸附测定(ELISA)筛选所得杂交瘤的抗体产生。
■ELISA结果表明,所有三种mAb均表现出对MERS-CoVS1抗原的强反应性。同样,dot-ELISA揭示了它们特异性识别病毒成分的能力,表明其诊断应用的潜力。在非变性条件下,Western印迹显示mAb对116KDa的特定条带具有强烈的反应性,对应于推定的MERS-CoVS1抗原。然而,在变性条件下没有观察到反应带,这表明抗体识别构象表位。中和测定显示对MERS-CoV没有体外反应性。
■这项研究使用杂交瘤技术成功地产生了三种针对MERS-CoV的小鼠单克隆抗体。使用ELISA和斑点ELISA测定,抗体表现出针对MERS-CoV抗原的强反应性。一起来看,这些发现强调了这些mAb作为MERS-CoV研究和诊断(社区和现场监测以及病毒抗原检测)的有价值工具的潜在用途.
UNASSIGNED: Middle East Respiratory Syndrome Coronavirus is a highly pathogenic virus that poses a significant threat to public health.
UNASSIGNED: The purpose of this study is to develop and characterize novel mouse monoclonal antibodies targeting the spike protein S1 subunit of the Middle East Respiratory Syndrome Corona Virus (MERS-CoV).
UNASSIGNED: In this study, three mouse monoclonal antibodies (mAbs) against MERS-CoV were generated and characterized using hybridoma technology. The mAbs were evaluated for their reactivity and neutralization activity. The mAbs were generated through hybridoma technology by the fusion of myeloma cells and spleen cells from MERS-CoV-S1 immunized mice. The resulting
hybridomas were screened for antibody production using enzyme-linked immunosorbent assays (ELISA).
UNASSIGNED: ELISA results demonstrated that all three mAbs exhibited strong reactivity against the MERS-CoV S1-antigen. Similarly, dot-ELISA revealed their ability to specifically recognize viral components, indicating their potential for diagnostic applications. Under non-denaturing conditions, Western blot showed the mAbs to have robust reactivity against a specific band at 116 KDa, corresponding to a putative MERS-CoV S1-antigen. However, no reactive bands were observed under denaturing conditions, suggesting that the antibodies recognize conformational epitopes. The neutralization assay showed no in vitro reactivity against MERS-CoV.
UNASSIGNED: This study successfully generated three mouse monoclonal antibodies against MERS-CoV using hybridoma technology. The antibodies exhibited strong reactivity against MERS-CoV antigens using ELISA and dot ELISA assays. Taken together, these findings highlight the significance of these mAbs for potential use as valuable tools for MERS-CoV research and diagnosis (community and field-based surveillance and viral antigen detection).