鸭源新城疫病毒HN蛋白单克隆抗体的制备及与不同分离株的反应性

Preparation of Monoclonal Antibodies against HN Protein of NDV Isolated from Duck and their Reactions with Different NDV Strains

为了制备鸭源新城疫病毒基因Ⅶ型毒株HN蛋白的单克隆抗体,将构建的原核表达载体p ET28(a)-HN转化BL21(DE3)进行原核表达,纯化HN重组蛋白,并免疫6周龄BALB/c小鼠,分离脾淋巴细胞与SP2/0细胞进行融合,克隆培养并筛选杂交瘤细胞株,制备单克隆抗体,并对其生物学特性进行了鉴定。结果显示,表达纯化的重组HN蛋白分子质量约为49 ku,用ELISA方法成功筛选出4株针对新城疫Ⅶ型毒株HN蛋白的单克隆抗体,其中2H7抗体抗原谱较广,具有中和活性,为进一步研究HN蛋白功能和临床诊断奠定了基础。

The recombinant plasmid pET28 （a） - HN was transformed into BL21 （ DE3 ） to purify the recombinant HN protein. The 6 - week - old BALB/c mice were immunized by the protein. The spleen lymphocytes were separated and conducted the cell fusion with SP2/0 cells at 3 days after the last immunization. The hybridoma cell lines were obtained by clone culture and indirect ELISA assay. Then the monoclonal antibodies against HN protein of newcastle disease virus （NDV） were preparated. Their biologic properties were identi- fied. The results showed that the molecular mass of purified recombinant HN protein was about 49 ku. Four monoclonal antibodies were screened. In which, the antibody 2H7 had broad antigen spectrum and neutralization activity. This study provided a basis for further research on the HN protein function and clinical diagnosis.