猪瘟病毒单克隆抗体的制备及其免疫组织化学研究

Preparation and immunohistochemical studies of monoclonal antibody against CSFV

通过杂交瘤技术制备猪瘟病毒单克隆抗体,对酶联免疫吸附试验筛选的阳性单克隆抗体进行免疫组织化学研究,从中发现1株单克隆抗体能与感染猪瘟病毒的猪肾、肝病理组织切片及PK15细胞产生特异性染色反应,与非感染猪的肾、肝组织切片和PK15细胞无染色反应。单克隆抗体与猪瘟病毒感染的PK15细胞或猪肝、肾小血管内皮细胞中的猪瘟病毒呈现较强的特异性棕色着染,表明该单抗是针对猪瘟病毒的单克隆抗体,对研究猪瘟病毒在宿主细胞中的生命活动和繁殖定位有非常重要的意义。该单抗命名为YNF,培养上清液和腹水的抗体效价分别为1∶80和1∶10 000。抗体蛋白属IgG类,亚类为IgG2/κ。

A strain of monoclonal antibody（McAb） against classical swine fever virus（CSFV） was prepared by the hybridoma technique. The McAb, named YNF with isotype of IgG2/κ in mouse IgG subclass, showed specific reaction to the CSFV-infected PK15 cells, the CSFV-infected tissues of kidney and liver of pigs, but showed no response to the PK15 cells and the tissues of kidney and liver of pigs, which were not infected with CSFV. The ELISA titres of the McAb were 1 ： 80 and 1 ： 10 000 in the supernatant of hybridoma cell and mouse ascites, respectively. The McAb performed a bright immunostaining to CSFV in PK15 cells and endotheliocytes of small vessels in the tissues. These results demonstrated that the McAb is a specific reagent for detection of the CSFV distribution in the host.