CAJ | 학술논문

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E2 MAb的杂交瘤细胞;测定MAb亚型并纯化后.间接ELJSA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1:25 600.
위제비항저온병독(CSFV)단극륭항체(MAb),본실험이표체CSFV E2낭막당단백적수포구염가병독면역BALB/c소서,취기비세포여골수류세포SP2/0진행융합;이용간접ELISA방법화휴대형광소매(Luciferase)보고기인적HIV-luc/CSFV-E1&E2가병독계통사선분비중화성E2 MAb적잡교류세포;측정MAb아형병순화후.간접ELJSA방법측정MAb적효개;채용western blot감정MAb적특이성친화력;이용HIV-luc/CSFV-E1&E2가병독진행체외중화시험,분석MAb억제병독감염적능력.결과표명본실험획득료1주분비중화성MAb적잡교류세포9C8,해MAb능여E2단백특이성결합,이차체외억제시험중화효개대우1:25 600.
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 bybridoma cells were screened by indirect ELISA and retroviral pseudotype virus HTV-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.