鹌鹑减蛋综合征病毒单克隆抗体的制备及生物学特性

以鹌鹑减蛋综合征QAV C94病毒为抗原免疫BALB/c小鼠 ,制备单克隆抗体并对其生物学特性进行鉴定。获得 5株分泌单克隆抗体的细胞株 (命名为H1、H3、H4、H6和H8)。这 5株杂交瘤细胞可长期稳定地分泌单克隆抗体 ,分泌的单克隆抗体为IgG2a亚类 ,具有高度的特异性 ,只特异地作用于鹌鹑减蛋综合征病毒 ;微量血凝抑制试验显示H4有血凝抑制活性 ,腹水效价为 1∶2 6 ;这 5株单克隆抗体都没有沉淀活性 ,也不与CELO抗原发生沉淀反应 ;它们的亲和力以H4最大 ,相对亲和力依次为H4 >H6 >H1 >H8>H3。

Preparation and biological characterization of monoclonal antibodies against egg drop syndrome virus from quail

BALB/c mice were immunized with chromatographically purified egg drop syndrome virus QAV C94 strain from quail, and splenocytes were prepared three days after the final booster immunization. Fusion of lymphocyte and myeloma SP2/0 cell line was carried out with the fusogen 50% polyethylene glycol. Five antibody secreting cell lines, named H1,H3,H4,H6 and H8, were finally obtained after selective cultivation, specific antibody detection and cloning. It was shown that the hybridoma cell lines can secrete antibody stably for a long time, the antibodies were belong to IgG2a. Biological properties showed that all antibodies were highly specific to QAV C94 virus. H4 monoclonal antibody had the activity of hemagglutination inhibition. The HI titers of ascitic fluids were 2 6 . None of these McAbs had immunoprecipitation capacity. Their affinity, sorted as H4>H6>H1>H8>H3, were calculated with optical densities determined by indirect ELISA.