摘要
为制备抗猪瘟病毒(CSFV)单克隆抗体(MAb),本实验以表达CSFV E2囊膜糖蛋白的水泡口炎假病毒免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞SP2/0进行融合;利用间接ELISA方法和携带荧光素酶(Luciferase)报告基因的HIV-luc/CSFV-E1&E2假病毒系统筛选分泌中和性E2MAb的杂交瘤细胞;测定MAb亚型并纯化后,间接ELISA方法测定MAb的效价;采用western blot鉴定MAb的特异性亲和力;利用HIV-luc/CSFV-E1&E2假病毒进行体外中和试验,分析MAb抑制病毒感染的能力。结果表明本实验获得了1株分泌中和性MAb的杂交瘤细胞9C8,该MAb能与E2蛋白特异性结合,而且体外抑制试验中和效价大于1∶25600。
To prepare E2 neutralizing monoclonal antibody（MAb） against classical swine fever virus（CSFV） ,BALB/c mice were immunized with recombinant vesicular stomatitis virus（rVSV） expressing the envelope glycoprotein E2 of CSFV;mouse splenic cells were fused with SP2/0 cells and hybridoma cells were screened by indirect ELISA and retroviral pseudotype virus HIV-luc/CSFV bearing CSFV E1 and E2 glycoproteins. The specificity and the antigen-binding activity of the MAb were identified by indirect ELISA and western blot,and neutralizing activity against CSFV was measured as inhibitions of luciferase reporter gene by pseudotype neutralization assay. One hybridoma（9C8） secreting neutralizing MAb against CSFV was successfully obtained;The MAb showed specificity against the E2 protein and had a neutralization titre more than 1∶25 600.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2010年第4期289-293,共5页
Chinese Journal of Preventive Veterinary Medicine
关键词
猪瘟病毒
中和性单克隆抗体
假病毒
中和试验
CSFV
neutralizing monocolonal antibody
pseudotyped virus
neutralization assay
