抗狂犬病病毒CVS-11株单克隆抗体的制备及鉴定  

Preparation and Characterization of monoclonal antibodies to rabies virus CVS-11 strain

作　　者：曹蕾[1] 李雄[2] 张守峰[3] 李六斤[1] 扈荣良[3] 唐青[1] 

机构地区：[1]第四军医大学实验动物研究中心,西安710032 [2]新疆维吾尔自治区疾病预防控制中心寄生虫病科 [3]解放军军事医学科学院军事兽医研究所

出　　处：《中华实验和临床病毒学杂志》2009年第1期68-70,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：国家863项目（SQ2006AA02Z112882）;国家自然科学基金重点资助项目（30630049）;公益性行业（农业）科研专项（200803014）

摘　　要：目的制备抗狂犬病毒单克隆抗体，为建立快速准确的狂犬病毒抗原检测方法奠定基础。方法将狂犬病病毒CVS-11株纯化浓缩后免疫6～8周龄雌性BALB／c小鼠，取其脾细胞与SP2／0骨髓瘤细胞进行融合，经细胞克隆和间接ELISA筛选，获得稳定分泌抗狂犬病毒单克隆抗体的杂交瘤细胞株。小鼠腹腔注射法制备大量单克隆抗体并测定腹水效价，用G蛋白亲和层析柱进行纯化，间接ELISA法和间接荧光法鉴定单克隆抗体的类型、特异性及敏感性。结果细胞融合率达100％，经克隆筛选获4株稳定分泌抗狂犬病病毒抗体的杂交瘤细胞株，其腹水效价分别为1×10^4，1×10^5，1×10^4和1×10^5；4株单抗均为IgG类型且特异性好。结论制备的单抗具有良好特异性和敏感性。Objective To get the high specific and sensitive mononclonal antibodies against RV. Methods The myeloma cell line SP2/0 fused with the spleen cell of 6 - 8 weeks old BALB/c mice immunized with the CVS-11 virus antigen. The hybridized fusing cells were chosen by indirect ELISA detection and the positive hybridizing cells were amplified through mouse abdomen injection and the mouse McAbs ascites was purified by Protein A Sepharose 4 Fast Flow（ Pharulacia Company） . The specificity and sensitivity of the McAbs was identified by indirect ELISA and indirect DFA test. Results The cell fusion rate reachs 100% and the indirect ELISA results showed that the McAbs ascites titer were 1×10^4, 1 ×10^5, 1 ×10^4 and 1 ×10^5;The immunoglobulin G type McAbs show no cross reaction with other related viruses. Conclusion The high specific and sensitive mononclonal antibodies of RV can be used for rapid RV diagnosis.

关 键 词：狂犬病病毒 抗体 单克隆 检测方法 

分 类 号：R3[医药卫生—基础医学]