牛γ干扰素单克隆抗体的制备与抗原捕获ELISA检测方法的建立  

Development of monoclonal antibodies against Bovine gamma-IFN and establishment of antigen capture ELISA for bovine gamma-IFN detection

作　　者：张月美[1] 陈伟业[1] 葛金英[1] 杨帆[2] 步志高[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部兽医公共卫生重点开放实验室,黑龙江哈尔滨150001 [2]东北农业大学,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2009年第12期953-957,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家"攻关"项目(2004BA519A19;2005BA711A10);国家"973"项目(2005CB523200)

摘　　要：为了建立一种简便、快速、可靠和经济的牛γ干扰素(BoIFN-γ)的免疫学检测方法,本研究通过杂交瘤技术建立了2株抗rBoIFN-γ单克隆抗体(MAb)2G6和5F9。Western blot、抗病毒活性阻断实验表明2株MAb与rBoIFN-γ有高亲和活性。相加ELISA表明2G6和5F9识别rBoIFN-γ不同的抗原表位。利用2G6作为捕获抗体,5F9-HRP作为检测抗体建立的抗原捕获ELISA方法可特异性的检测不同系统表达的rBoIFN-γ,而且与rBoIFN-α、rBoIFN-β不发生交叉反应。该方法的建立为抗原捕获ELISA定量检测BoIFN-γ试剂盒的研制奠定了基础。An antigen capture ELISA was established by utilizing high affinity monoclonal antibodies （MAb） against BoIFN-γ. Two hybridomas 2G6 and 5F9 were developed and confirmed by western blot and antiviral blocking assay. The additive ELISA demonstrated that the two MAbs recognized different antigenic regions. Antigen capture ELISA based on these MAbs is highly specific and sensitive, proving a quick and reliable method for detecting BoIFN-γ.

关 键 词：BoIFN-γ 单克隆抗体 ELISA 检测 

分 类 号：S852.42[农业科学—基础兽医学]