摘要
利用IPTG诱导大肠杆菌表达牛-γ干扰素融合蛋白(GST-BovIFN-γ),经聚丙烯酰胺凝胶电泳(SDS-PAGE)后,切取融合蛋白条带作为抗原免疫BALB/c小鼠。利用杂交瘤技术研制抗牛-γ干扰素(BovIFN-γ)的特异性单克隆抗体(mAb)。采用间接酶联免疫吸附试验(ELISA)方法筛选、Western-blotting、间接免疫荧光(IFA)方法进行鉴定,筛选出2株分泌特异性抗BovIFN-γ的杂交瘤细胞株,分别命名为BovIFN--γ4A3和BovIFN--γ4G5。该单抗可以与重组GST-BovIFN-γ和真核细胞表达的rBovIFN-γ反应,但不能与GST和鸡IFN-γ反应,说明单抗具有很好的特异性。进一步研究表明单抗能中和BovIFN-γ对水泡性口炎病毒(VSV)对牛肾细胞的致病变作用。这些单抗可用于奶牛结核病的监测和防治。
Two hybridoma cells secreting monoclonal antibodies against bovine γ-interferon were obtained by fusion between SP2/0 cells and spleen cells from mice immunized with GST-BovlFN-γ fusion protein expressed on IPTG-induced E. coll. This fusion protein was recovered from gel of SDS-PAGE, and used as immunogen to immunize BALB/c mice. The monoclonal antibodies thus obtained were designated as BovlFN-γ4A3 and BovlFN-γ4G5 respectively. As demonstrated by ELISA and IFA, these monoclonal antibodies reacted only with BovIFN-γ, but not with GST or chicken IFN-γ. In addition, they could neutralize the cytopathic effect on bovine kedney cells induced by vesicular Stomatitis virus （VSV） and showed high specificity and stability as demonstrated in indirect ELISA, Western blotting and IFA assays. It is evident that these monoclonal antibodies can be used for the surveillance and control of bovine tuberculosis in milk cows.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2006年第8期755-758,共4页
Chinese Journal of Zoonoses
基金
江苏省"333"工程人才培养基金(2002)资助
关键词
奶牛
Γ-干扰素
单克隆抗体
bovine
interferon-gamma
monoclonal antibody
