猪繁殖与呼吸综合征病毒GP5蛋白中和性单克隆抗体研制
evelopment and Biological Characteristics of Monoclonal Antibodies against GP5 protein of Porcine Reproduction and Respiratory Syndrome Virus
目的 研制猪繁殖与呼吸综合征病毒GP5蛋白的单克隆抗体(McAb)。 方法 将GP5基因克隆入真核表达载体pcDNA3.1，利用构建成的重组真核质粒免疫BALB / c小鼠, 采用杂交瘤技术制备抗PRRSV GP5蛋白单抗。建立间接ELISA方法筛选阳性克隆。并对杂交瘤细胞株进行鉴定，分析单抗特性。 结果 获得3株可分泌特异性抗PRRSV GP5蛋白抗体的杂交瘤细胞株，分别命名为4C9、5D10、5F12。其中5D10的Ig亚类为IgM。ELISA检测杂交瘤细胞培养上清抗体效价为1：64～1：1024,腹水效价为1：3200~1：10240, 同时用Western blot、IFA和IPMA检测，结果均为阳性。5D10和5F12的相对亲和力不同，证明其识别不同的抗原位点。5D10、5F12具有病毒中和活性，中和效价分别为1：40和1：80。 结论 获得2株抗PRRSV GP5蛋白的单克隆抗体,为进一步对PRRSV诊断和免疫预防研究奠定了重要基础。
o prepare monoclonal antibodies（McAb） against GP5 protein of porcine reproduction and respiratory syndrome virus . The plasmids encoding GP5 genes of PRRSV-S1 strain was used as antigen. Balb/ c mice were immunized by subcutaneous injection. The induced spleen cells were fused with SP2/ 0 mydoma cell . The McAbs anti GP5 were obtained by screening with indirect ELISA and subcloned by 4 times, and were preliminary characterized. Three hybridoma cell strains steadily secreting monoclonal antibodies(McAb) against GP5 protein of PRRSV were obtained, which were separately named4C9、5D10 and 5F12。The ascites titers of PRRSV-specific ELISA antibodies obtained by mice were be 1 ∶3 200 to 1 ∶10240. Relative affinity of 5D10 were tighter than that of 5F12.It indicated that these McAbs were against different regions of GP5 protein of PRRSV .More over, the McAbs 5D10 and 5F12 had PRRSV-specific neutralization activity. The titers of the neutralizing antibodies were 1:40 and 1:80. The isotype of the 5D10 was IgM .It should be very useful to diagnose and prevent PRRSV infection.
马苏、李玉峰 、姜平、段舒怡
基础医学生物科学研究方法、生物科学研究技术微生物学
猪繁殖与呼吸综合征病毒GP5蛋白质粒单克隆抗体、中和抗体
PRRSVMonoclonal antibodyGP5 proteinNeutralization antibody
马苏,李玉峰 ,姜平,段舒怡.猪繁殖与呼吸综合征病毒GP5蛋白中和性单克隆抗体研制[EB/OL].(2006-11-13)[2025-06-12].http://www.paper.edu.cn/releasepaper/content/200611-324.点此复制
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