抗鸡白细胞介素4单克隆抗体的制备及其鉴定

发布时间：2018-02-04 12:34

本文关键词： 鸡白细胞介素(ChIL-) 单克隆抗体　出处：《细胞与分子免疫学杂志》2017年07期 　论文类型：期刊论文

【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)×104;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。
[Abstract]:Objective to prepare and identify monoclonal antibody against chicken interleukin-4 (ChIL-4). Methods Recombinant chicken IL-4 GST-ChIL-4 was prepared and identified. To immunize 8-week-old BALB/c mice as an immunogen. Using recombinant protein His-ChIL-4 as detection antigen, indirect Elisa was established to screen positive hybridoma cell lines. ChIL-4 mAb cell lines secreted stably and stably were prepared by limited dilution method. The specificity of mAb was analyzed by indirect Elisa, indirect immunofluorescence assay (IFA) and Western blot. Results four mAb strains resistant to ChIL-4 were obtained. The ELISA titer of ascites mAb was 1.6 ~ 6.4 脳 10 ~ (4) 脳 10 ~ (4), respectively, and it was named as 6A8A ~ (18) F _ (12) ~ (19) F _ (1) ~ (20) G _ (3). The results of ELISAA IFA and Western blot showed that the four mAb strains were mAb-specific to recombinant ChIL-4 and did not recognize recombinant bovine IL-4. The ability of other proteins such as chicken interferon 纬 (IFN- 纬). Conclusion the anti-mAbs of ChIL-4 were successfully prepared.
【作者单位】：扬州大学江苏省人兽共患病学重点实验室江苏省动物重要疫病与人兽共患病防控协同创新中心医学院;
【基金】：国家自然科学基金(30972171,81472815)
【分类号】：S852.4
【正文快照】： 白细胞介素4(interleukin 4,IL-4)主要由2型辅助T(T helper type 2,Th2)细胞分泌,具有刺激Th2细胞分化、增殖的能力,是Th2细胞免疫应答的重要调节因子[1-2],并对滤泡辅助T(follicular helperT cell,Tfh)细胞、B细胞、单核细胞、巨噬细胞等其他多种细胞均具有免疫调节功能,从而

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