摘要
为了制备高质量高效价抗牛γ干扰素(Bo IFN-γ)的单克隆抗体,本研究将Bo IFN-γ基因克隆到His标签的大肠杆菌表达载体p ET32a+中,诱导表达并纯化蛋白作为抗原免疫BALB/c小鼠制备单克隆抗体。结果筛选到9株单抗,分别命名为Bo IFN-γ-1E10、Bo IFN-γ-3C2、Bo IFN-γ-3E1、Bo IFN-γ-6C6、Bo IFN-γ-6H9、Bo IFN-γ-6B6、Bo IFN-γ-6E5、Bo IFN-γ-2A10和Bo IFN-γ-2G10。ELISA、Western blot和间接免疫荧光试验结果显示,制备的单克隆抗体与原核、真核表达的Bo IFN-γ都具有良好的反应性。本研究制备获得的针对牛γ干扰素的单克隆抗体为建立检测天然Bo IFN-γ的方法提供了重要的生物材料。
To generate monoclonal antibodies against bovine IFN- γ（ Bo IFN- γ）,the fragment of Bo IFN- γ in the p GEX- 6P- 1 vector was sbucloned into p ET32 a vector. The purified His- Bo IFN- γ fusion protein was injected into BALB / c mice as immunogen. Nine positive monoclonal antibodies,named as Bo IFN- γ- 1E10,Bo IFN- γ- 3C2,Bo IFN- γ- 3E1,Bo IFN- γ- 6C6,Bo IFN- γ- 6H9,Bo IFN- γ- 6B6,Bo IFN- γ- 6E5,Bo IFN- γ- 2A10 and Bo IFN- γ- 2G10,were obtained. The results of ELISA,Western blot and immunofluorescence assay（ IFA） demonstrated that these monoclonal antibodies were able to recognize not only prokaryotic but also eukaryotic expression Bo IFN- γ protein. The generated monoclonal antibodies in this study will provide important reagents for further establishment of a sandwich ELISA for the detection of natural Bo IFN- γ.
出处
《畜牧与兽医》
北大核心
2016年第9期11-15,共5页
Animal Husbandry & Veterinary Medicine
基金
江苏省科技支撑计划(BE2013391)
江苏省优势学科项目
关键词
牛γ干扰素
原核表达
单克隆抗体
Bo IFN-γ
prokaryotic expression
monoclonal antibody
