重组马γ-干扰素抗病毒活性测定及单克隆抗体抑制活性鉴定  

Antiviral Activity Determination of Recombinant Equine Interferon-gamma and Identification Inhibited Antiviral Activity of Monoclonal Antibodies

作　　者：白宇[1] 陈伟业[1] 童铁钢[1] 张维军[1] 徐树兰[1] 王群[1] 孙庆歌[1] 刘光亮[1] 步志高[1] 吴东来[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,哈尔滨150001

出　　处：《生物工程学报》2008年第7期1258-1262,共5页Chinese Journal of Biotechnology

基　　金：国家高科技发展计划(863)项目(No.2006AA10A203);黑龙江省自然科学基金重点项目(No.ZJN-0602-01)资助~~

摘　　要：为了测定大肠杆菌和杆状病毒表达的重组马γ-干扰素是否具有抗病毒活性,利用这两种干扰素处理马胎肾细胞(EFK-78),然后接种表达绿色荧光蛋白(GFP)的重组水泡性口炎病毒(VSV*GFP),观察干扰素对病毒表达GFP的抑制,测出其抗病毒活性单位分别为1×103AU/mL、1×105AU/mL。评价了制备的九株抗重组马γ-干扰素单克隆抗体是否可抑制重组马γ-干扰素抗病毒活性,证实其中一株可中和重组马γ-干扰素的抗病毒活性。结果表明:杆状病毒表达的马γ-干扰素具有较高的抗病毒活性,其活性可被一株制备的抗重组马γ-干扰素单克隆抗体抑制;首次获得原核表达的具有抗病毒活性的马γ-干扰素。Equine interferon-gamma （eIFN-γ） expressed both in E. coli and baculovirus were evaluated for antiviral activity against recombinant Vesicular Stomatits Virus expressing green fluorescence protein （rVSV-GFP） in EFK-78 cells. The assays were conducted in 96-well plate. Virus infectivity was measured by quantifying GFP-positive cells, instead of quantifying the CPE reduction. Prior to infection of EFK-78 cells with rVSV-GFP, the cells were incubated with eIFN-γ. The GFP expression in the EFK-78 cells dramatically decreased in the cells treated with eIFN-γ in a dose-dependent manner, comparing with the mock-treated cells. The titers of antiviral activity were 1 × 10^3AU/mL and 1 × 10^5AU/mL of eIFN-Texpressed from E. coli and baculovirus, respectively. The antiviral activities of the recombinant eIFN-γ were highly efficient and specific, as it was blocked by mAbs against eIFN-γ.

关 键 词：马γ-干扰素 抗病毒活性测定 单克隆抗体 

分 类 号：Q78[生物学—分子生物学] S852.43[农业科学—基础兽医学]