马立克氏病病毒囊膜糖蛋白B基因原核细胞表达产物单克隆抗体的制备

Preparation of Monoclonal Antibody of Marek′s Disease Virus(MDV) Glycoprotein B Expressed in E.coli

用大肠杆菌 BL2 1表达了马立克氏病病毒 (MDV)强毒 GA株的囊膜糖蛋白 B(g B)基因 ,通过 SDS- PAGE电泳分离表达蛋白条带 ,切下并碾碎后作为免疫原制备单克隆抗体。通过 EL ISA和间接免疫荧光试验 (IFA) ,得到 1株阳性单克隆抗体杂交瘤细胞株 7C8。该单抗腹水的 EL ISA效价为 1∶ 2 1 2 ,IFA效价为 1∶ 80 0 ,与 MDV不同致病型毒株CVI988、GA、RB1B、MD11、6 48A株感染的鸡胚成纤维细胞 (CEF)均呈 IFA阳性。1∶ 10 0稀释时与 GA株感染的 CEF在斑点酶联免疫吸附试验 (dot- EL ISA)中呈阳性。

Glycoprotein B gene(gB) of virulent MDV GA strain expressed in E.coli was cut off from SDS-PAGE,then inoculated it to BALB/c mice directly to prepare monoclonal antibody.1 strain of monoclonal hybridoma cells 7C8 was obtained by ELISA and Indirect Fluorescence Assay(IFA).Its ascites fluids titre of ELISA and IFA is 1∶2 12 and 1∶800 respectively.The McAb can also identify CEF infected by MDV through dot-ELISA.