本研究以原核表达、纯化的扎伊尔株埃博拉病毒(Ebola virus,EBOV)VP40蛋白片段免疫BALB/c小鼠,通过杂交瘤技术制备单克隆抗体,获得一株分泌抗EBOVVP40蛋白单克隆抗体的杂交瘤细胞系。以重组杆状病毒表达EBOV VP40蛋白为抗原,经Western blot和间接免疫荧光检测该单克隆抗体显示出良好的特异性免疫反应。采用该株杂交瘤细胞系经腹腔注射接种8日龄BALB/c小鼠,制备腹水,并经纯化和HRP标记,获得HRP标记的抗EBOV VP40蛋白单克隆抗体。从而,初步建立一种抗原捕捉ELSIA诊断方法,以原核表达EBOV VP40蛋白片段兔抗血清为一抗,应用HRP标记的抗EBOV VP40蛋白单克隆抗体检测杆状病毒表达的重组EBOV VP40蛋白显示出良好的敏感性和特异性。结果表明,初步建立的抗原捕捉ELSIA诊断方法有希望成为一种理想的检测EBOV感染的诊断方法。
In this study, a monoclonal antibody (mAbs) against VP40 protein of Ebola virus (EBOV), Zaire species, were produced using a recombinant VP40 derived from E. coli. The mAbs was shown to be highly specific in Western blot and indirect immunofluorescence detection. An antigen capture ELSIA was developed using mAbs purified from ascites and labeled with the horseradish peroxidase (HRP). The assay was able to detect protein VP40 of EBOV expressed by recombinant baculovirus. The established enzyme-linked immunosorbent assay therefore could be a useful diagnosis tool for use in the field.