抗牙鲆淋巴囊肿病毒单克隆抗体的制备

用本实验室分离纯化的淋巴囊肿病毒(Lymphosystis Disease Virus China，LCDVcn)作为抗原，感染BALB／c小鼠，取被免疫小鼠的脾细胞与骨髓瘤细胞(Sp2／0融合；ELISA法检测、筛选阳性克隆，经过3次克隆化，首次成功获得18株抗牙鲆淋巴囊肿病毒的单克隆抗体。

Production and Characterization of Monoclonal Antibodies against Paralichthys Olivaceus Lymphosystis Disease Virus

By using hybridoma-monoclonal antibody technology, 18 hybridoma cell lines which secrete monoclonal antibodies(Mab)against Lymphosystis Disease Virus(LCDV)had been established. And the characteristics of these Mabs had been described. In isotyping analysis, the results showed that among these Mabs, 12 of them are IgG 2b , 6 of them are IgM; all of these Mabs have the ability to bind LCDV by ELISA and their titer ranged from 10 4 to 10 7. Further experiments proved that all of them only specifically bound to LCDV, and did not have any cross reaction with the hemorrhagic fever renal syndrome virus(HFRSV), Japanese encephalitis virus (JEV)or herpes virus. The measuring sensitivity of the Mab YpH4A3 to purified LCDV was as low as 35ng. All of these results demonstrated that Mabs were highly specific and sensitive to LCDV, and had the great potential to be used for pathogen diagnosis. Furthermore, we can find out if some of the Mabs are neutralizing antibodies. Then we can use the neutralizing antibodies to test the protective antigen sequence of LCDV and produce the polypeptide vaccine against LCDV.