鸭星状病毒单克隆抗体的制备及鉴定

Preparation andidentification of Monoclonal Antibodies against Duck Astrovirus

为了制备针对鸭星状病毒(Duck astrovirus,DAstV)衣壳蛋白的单克隆抗体,以DAstV C-NGB株为免疫原,用ORF2重组蛋白作为抗原进行筛选,获得7株稳定分泌抗DAstV ORF2蛋白单抗的杂交瘤细胞。取E5C1株和C10F6株进行鉴定,结果表明,2株单抗均为IgM,轻链均为κ型;细胞上清和小鼠腹水的效价分别为10~5和10~6以上;免疫印迹试验表明,2株单抗均能识别重组ORF2蛋白;特异性试验表明,E5C1株杂交瘤细胞的培养上清只与DAstV反应。

For preparing monoelonal antibodies （Abs） against duck astrovirus （DAstV） capsid protein, the DAstV C -NGB isolate was used as immunogen and the purified recombinant ORF2 protein as antigen to secreen specific antibodies. Seven hybridoma cell lines that stably secreted Abs against DAstV ORF2 were obtained. Two selected Abs, namely E5C1 and C10F6, were identified as IgM with light chains belonging to K type. The titers of both Abs in cell culture supematants and ascites were determined to be 1 ： 105 and 1 ： 106 , respectively. The reactions of the E5C1 and C10F6 Abs with the recombinant ORP2 protein were confirmed by Western Blotting. The E5C1Ah in culture supematants was tested to react only with DAstV in specificity tests.