新城疫病毒F48E9株重组NP蛋白单克隆抗体制备及鉴定

摘要

Newcastle disease virus (NDV)was prepared in order to lay the foundation for NDV antigen epitope research and establishment of detection methods. The resnlt showed that BALB/c mice were immunized with pET32a-NP recombinant protein of NDV F48E9, two hybridoma cell lines steadily secreting McAb against NP protein of NDV were obtained by hybridomatechnology and indirect ELISA, and the cell lines were named 1G3 and 4B12. Their titers of ascites were 1:2.56×105 and 1:5.12 ×105, respectively, by indirect ELISA. The isotypes of McAbs were all IgG1. They were highly specific to recombinant NP protein by Western blot. It showed yellow and green fluorescence that cells infected with NDV by Indirect immunofluorescence assay (IFA). Additive ELISA showed that the two McAbs could recognize different epitopes.%制备新城疫病毒(NDV)NP蛋白单抗,以期为NDV抗原表位研究及检测方法建立方面奠定基础.利用NDV F48E9,株pET32a-NP重组蛋白免疫BALB/c小鼠,采用杂交瘤细胞技术,问接ELISA筛选,获得了2株稳定分泌抗NP重组蛋白单克隆抗体的杂交瘤细胞株,分别命名为1G3、4B12.经间接ELISA测定,腹水抗体效价分别为1∶2.56x 105、1∶5.12x105.亚类鉴定结果表明这两株单抗均为IgG1.Western blot分析结果显示,1G3、4B12均能特异性识别重组NP蛋白.与NDV感染细胞经间接免疫荧光试验检测均呈黄绿色荧光.经相加ELISA测定表明两株单抗识别的抗原表位不同.