抗乙型脑炎病毒非结构蛋白NS1单克隆抗体的制备与鉴定

Production and characterization of the monoclonal antibodies against NS1 protein of Japanese encephalitis virus

本研究利用原核表达的乙型脑炎病毒(JEV)SA14-14-2株非结构蛋白NS1作为免疫原,免疫8周龄BALB/c小鼠,采用淋巴细胞杂交瘤技术进行融合,共获得4株特异性针对JEV NS1的杂交瘤细胞,分别命名为1H6、2C3、3A7、4C8,经测定1H6单抗亚类属于IgG2b,其他3株为IgG1,轻链均为κ链。4株杂交瘤细胞诱生小鼠腹水效价分别达1∶20480、1∶2560、1∶20480、1∶10240,western blot证实所得杂交瘤细胞分泌的抗体均可与JEV NS1蛋白发生特异性反应,间接免疫荧光试验表明1H6、3A7、4C83株单抗能够识别天然的JEV NS1蛋白。本研究为进一步探究JEV NS1蛋白结构及其功能奠定了基础。

Four monoclonal antibodies （McAb） against Japanese encephalitis virus （JEV） nonstructuml protein （NSI） of SA14-14-2 strain were produced. These McAbs were generated by fusing BALB/c mice myeloma cell line SP2/0 with spleen lymphocytes from BALB/c mice immunized with prokaryotic expressed NS and were designated as 1H6, 2C3, 3A7 and 4C8, respectively. The McAb IH6 belonged to the subtype of IgG2b, while 2C3, 3A7 and 4C8 were identified as IgG1 subtype, and the light chains of all antibodies were n type. The antibody titer of ascites were 1：204 800, 1：25 600, 1：204 800 and 1：102 400 respectively. Western blot showed that the antibodies were specific to JEV NS1. Indirect immunofluorescence assay showed that 3 of 4 McAbs recognized JEV NSI protein in JEV infected BHK-21 cells. McAbs reported here may provide valuable tools for further investigation of the structure and function of JEV NS 1 protein.