禽脑脊髓炎病毒单克隆抗体的制备、鉴定与初步应用

为获得禽脑脊髓炎病毒(Avian Encephalomyelitis virus,AEV)VP1蛋白的单克隆抗体,通过原核表达AEV VP1蛋白,纯化后作为免疫原免疫BALB/c小鼠,并按常规方法制备杂交瘤细胞。经ELISA方法筛选阳性杂交瘤细胞,经过3次亚克隆获得2株杂交瘤细胞株,命名为4#、19#,并进行了抗体亚类的鉴定、Western-blot和IFA检测。结果显示：制备的株单克隆抗体亚型分别为IgG2b、IgG2a,Western-blot和IFA试验结果表明单克隆抗体均能与AEV发生特异性反应而与其他禽病常见病毒均无交叉反应。运用建立的IFA对单抗进行了初步运用,在外源病毒检测方面与经典方法符合率高。本研究成功制备了AEV单克隆抗体,为进一步建立AEV检测方法和深入研究AEV的生物学特性奠定了基础。

Preparation and Identification of Monoclonal Antibody against N Protein of Avian Encephalomyelitis Virus

In order to obtain the monoclonal antibody against VP1 protein of Avian encephalomyelitis virus, the VP1 protein of AEV was expressed in prokaryotic cells, purified and used as immunogen to immunize BALB/c mice, and the hybridoma cells were prepared according to the conventional method. Three hybridoma cell lines, named 4#, 19# were obtained by ELISA. The antibody subclasses were identified, Western-blot and IFA were detected. The results showed that the three monoclonal antibodies were IgG2b, IgG2a. Western-blot and IFA results showed that the monoclonal antibodies could specifically react with AEV and had no cross-reaction with other