抗鸡γ-干扰素单克隆抗体的研制及鉴定  

Preparation and characterization of monoclonal antibodies against chicken interferon-gamma

作　　者：戴华[1] 郑佳玉[1] 胡茂志[1] 陈俊华[1] 潘志明[1] 焦新安[1] 

机构地区：[1]扬州大学江苏省人兽共患病学重点实验室,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2007年第4期6-9,共4页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金资助项目(30425031);全国优秀博士学位论文作者专项基金资助项目(FANEDD200358)

摘　　要：应用淋巴细胞杂交瘤技术,以纯化的原核表达产物His-ChIFN-γ作为免疫原,以纯化的GST-ChIFN-γ作为检测原,制备抗鸡γ-干扰素(ChIFN-γ)mAb;采用ELISA、Dot-ELISA和ECL法鉴定单克隆抗体(mAb)的生物学特性。获得4株可稳定分泌抗ChIFN-γmAb的杂交瘤细胞株1G10、2C3、3E5、3E3,其腹水ELISA效价为2×104~8.2×105;Ig亚类分别为IgG1、IgG1、IgG1和IgG2a。Dot-ELISA和ECL试验结果均表明:4株mAb只与表达ChIFN-γ的重组大肠杆菌反应,而与未表达ChIFN-γ的菌株不反应,表明4株mAb均特异性针对ChIFN-γ。鸡脾脏淋巴细胞经PMA刺激后,经固定、破膜,用mAb 2C3标记,利用流式细胞术可成功地检测到表达ChIFN-γ的T淋巴细胞。mAb为家禽的免疫检测、免疫细胞功能分析和免疫调节等研究提供依据。Using lymphocyte hybridoma technique,8-week-old female BALB/c mice were immunized with purified His-ChIFN-γ expressed in the recombined bacteria,BL21(DE3)(pBCX-ChIFN-γ),then the spleen cells were collected and fused with Sp2/0-Ag-14 myeloma cells.Hybridoma cells were screened by indirect ELISA,the specificity of mAbs was characterized by indirect ELISA,Dot-ELISA and ECL.Four hybridoma cell lines secreting mAbs against ChIFN-γ were obtained and named as 1G10,2C3,3E5,3E3.The immunoglobulin subclasses of four ...

关 键 词：鸡 Γ-干扰素 单克隆抗体 流式细胞术 

分 类 号：S852.43[农业科学—基础兽医学]