鸡传染性支气管炎病毒特异性单克隆抗体的制备与鉴定

Preparation and Identification of Monoclonal Antibody against N Protein of Avian Infectious Bronchitis Virus

关键词:  鸡传染性支气管病毒  N蛋白  单克隆抗体

Key words:  Avian infectious bronchitis virus  N protein  monoclonal antibody

摘要:

为获得鸡传染性支气管炎病毒(Avian infectious bronchitis virus,IBV)N蛋白的单克隆抗体,通过原核表达IBV N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3株杂交瘤细胞株,命名为1#、18#、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3株单克隆抗体亚型均为IgG1,Western-blot和IFA试验结果表明单克隆抗体均能与IBV发生特异性反应而与其他禽病常见病毒均无交叉反应。本研究成功制备了IBV单克隆抗体,为进一步建立IBV检测方法和深入研究IBV的生物学特性奠定了基础。

Abstract:

In order to obtain the monoclonal antibody against N protein of Avian infectious bronchitis virus, the N protein of IBV 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 1#, 18# and 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 IgG1 subtype. Western-blot and IFA results showed that the monoclonal antibodies could specifically react with IBV and had no cross-reaction with other common avian viruses. In conclusion, the monoclonal antibody against N Protein of IBV was successfully prepared, which laid a foundation for further establishment of IBV detection methods and in-depth study of the biological characteristics of IBV.