中文摘要：

目的制备抗鸡白细胞介素4（ChIL-4）单克隆抗体（mAb）并进行鉴定。方法以重组鸡IL-4（GST-ChIL-4）作为免疫原免疫8周龄BALB/c小鼠,以重组蛋白His-ChIL-4作为检测抗原建立筛选阳性杂交瘤细胞株的间接ELISA;以有限稀释法制备稳定分泌抗ChIL-4 mAb细胞株;采用间接ELISA、间接免疫荧光法（IFA）以及Western blot法分析mAb的特异性。结果获得4株抗ChIL-4的mAb,分别命名为6A8、18F12、19F1、20G3,腹水mAb的ELISA效价为（1.6～6.4）×10^4;ELISA、IFA及Western blot结果显示该4株mAb均是特异针对重组ChIL-4的mAb,不具有识别重组牛IL-4、鸡γ干扰素（IFN-γ）等其他蛋白的能力。结论成功制备了抗ChIL-4的mAb。

英文摘要：

Objective To prepare and characterize the monoclonal antibodies（ m Abs） against chicken interleuikin-4（ ChIL-4）. Methods Using lymphocyte hybridoma technique,8-week-old BALB/c mice were immunized with purified recombinant ChIL-4（ r ChIL-4） with GST tag（ GST-ChIL-4）,and rChIL-4 with histidine tag（ His-ChIL-4） was utilized to set up an indirect ELISA to screen positive hybridomas. Limited dilution methods were used to generate hybridoma cell lines stably secreting m Abs against ChIL-4. The specificity of the m Abs was identified by indirect ELISA,immunofluorescent assay（ IFA） and Western blot. Results Four m Abs against r ChIL-4 were obtained and named 6A8,18F12,19F1 and 20G3. ELISA titers of the 4 m Abs were（ 1. 6 ～ 6. 4） × 10^4. ELISA,IFA and Western blot showed that the 4 m Abs could only react with rChIL-4,but not with the other proteins like bovine IL-4（ Bov IL-4）,chicken IFN-γ（ ChIFN-γ）. Conclusion The m Abs against r ChIL-4 were successfully obtained.