为制备鸭源新城疫病毒(NDV)的单克隆抗体(MAb),本研究以原核表达并纯化的鸭源NDV的核衣壳蛋白(yp)为免疫原,免疫6周龄~8周龄的BALB/c小鼠,通过淋巴细胞杂交瘤技术,经以重组蛋白为包被抗原的间接ELISA方法筛选,获得了2株能稳定分泌抗重组蛋白MAb的杂交瘤细胞株A1和B2,经ELISA检测A1株和B2株细胞上清效价均为2^12,诱生的腹水的效价分别为10^7和10^6。Westemblot和IFA检测显示.仅A1株能与鸭源NDVNP蛋白发生特异性反应。抗鸭源NDVNP蛋白MAb的研制,为鸭源NDV的免疫学诊断及致病机理的研究奠定基础。
To develop monoclonal antibodies (MAb) against duck-origin Newcastle disease virus. Eight-week-old female BALB/c mice were immunized with purified recombinant NP protein. Hybridoma cells were screened by indirect ELISA using recombinant NP protein as coating antigen. Two hybridoma cell lines stably secreting anti-NP protein, designated as A1 and B2 were scaned. Indirect ELISA results showed titers of both MAbs in cell culture medium were 212, while those of A1 and B2 MAbs in ascetic fluids were 107 and 106, respectively. Results of western blot and IFA indicated only MAb A1 could react with NP protein.