Department of General Surgery, The Affiliated Jiangsu Shengze Hospital of Nanjing Medical University, Suzhou, China.
Center for Public Health Research, Nanjing University, Nanjing, China.
J Med Virol. 2020 Aug;92(8):996-1006. doi: 10.1002/jmv.25615. Epub 2020 Feb 3.
The Ebola virus is highly infectious and characterized by hemorrhagic fever, headache, and so on with a high mortality rate. Currently, there are neither therapeutic drugs or vaccines against the Ebola virus nor fast diagnostic methods for the detection of Ebola virus infection. This study reported the induction and isolation of two monoclonal antibodies that specifically recognized the glycoprotein (GP) and secreted glycoprotein (sGP) of the Ebola virus. Plasmids encoding either GP or sGP were constructed and immunized BALB/c mice, accordingly purified sGP was boosted. The antisera were analyzed for binding activity against sGP protein in enzyme-linked immunosorbent assay (ELISA) and neutralization activity in a pseudotyped virus neutralization assay. A number of reactive clones were isolated and two monoclonal antibodies T231 and T242 were identified to react with both GP and sGP. Western blot and ELISA assays showed that the monoclonal antibodies could react with GP and sGP, respectively. Moreover, they could recognize Ebola pseudovirus by cellular immunochemistry assay. We labeled the monoclonal antibody T231 with biotin and analyzed the competitiveness of the two antibodies by the ELISA test. The results showed that the binding epitopes of the two monoclonal antibodies to sGP were partially overlapped. In summary, two GP-specific mAbs were identified, which will be used to detect the Ebola virus or investigate GP.
埃博拉病毒具有高度传染性，其特征为出血热、头痛等，死亡率高。目前，尚无针对埃博拉病毒的治疗药物或疫苗，也没有快速诊断埃博拉病毒感染的方法。本研究报告了两种单克隆抗体的诱导和分离，这两种单克隆抗体特异性识别埃博拉病毒的糖蛋白（GP）和分泌糖蛋白（sGP）。构建了分别编码 GP 或 sGP 的质粒，并免疫 BALB/c 小鼠，相应地对纯化的 sGP 进行了加强免疫。通过酶联免疫吸附试验（ELISA）分析抗血清与 sGP 蛋白的结合活性，通过假型病毒中和试验分析中和活性。分离出一些反应性克隆，并鉴定出两种单克隆抗体 T231 和 T242 可分别与 GP 和 sGP 反应。Western blot 和 ELISA 检测表明，这两种单克隆抗体可分别与 GP 和 sGP 反应。此外，它们还可以通过细胞免疫化学检测到埃博拉假病毒。我们用生物素标记单克隆抗体 T231，并通过 ELISA 试验分析了两种抗体的竞争能力。结果表明，两种单克隆抗体与 sGP 的结合表位部分重叠。总之，鉴定出了两种针对 GP 的特异性 mAb，可用于检测埃博拉病毒或研究 GP。
