| 摘要: |
| 以伪狂犬病毒(PRV)作为免疫原,免疫BALB/c小鼠,融合之后经间接ELISA、IFA和IHC试验进行杂交瘤筛选,共获得2株能分泌针对PRV单克隆抗体(简称单抗)的杂交瘤细胞株,分别命名为1E10和5B5,1E10亚类为IgG1,5B5亚类为IgG2b,单抗腹水的间接ELISA效价均达到1.0×108,染色体数目介于95~105之间。2株猪伪狂犬病毒单抗与猪细小病毒、猪繁殖与呼吸综合征病毒、猪圆环病毒2型、猪瘟病毒均无交叉反应,IFA结果显示单抗能与接种于猪睾丸细胞(ST)的PRV发生特异性反应,IHC结果显示单抗能用感染了PRV的猪神经组织发生反应,证实抗PRV单克隆抗体具有良好的特异性和敏感性,为猪伪狂犬病毒抗原表位分析及相关抗原抗体诊断试剂盒的研制奠定了基础。 |
| 关键词: 伪狂犬病毒 单克隆抗体 特异性 敏感性 |
| DOI： |
| 投稿时间：2014-01-21修订日期：2014-03-10 |
| 基金项目: |
|
| Preparation and Characterization of Monoclonal Antibody against Pseudorabies Virus |
|
| (Wu Han Chopper Biological Co.,Ltd.) |
| Abstract: |
| By taking PRV as immunogen, BALB/c mice as model and using indirect ELISA,IHC and IFA for hybridoma screening, we found out that 2 hybridoma cell line strains could secrete PRRSV monoclonal antibody (hereafter referred to as McAb), named1E10 and 5B5. 1E10 subgenera was for IgG1, and 5B5 subgenera for IgG2b, Single anti ascites of indirect ELISA valence achieved 1:1.0×107, The chromosome number is between 95~105. These two strains showed single resistance to porcine parvovirus, porcine pseudo rabies virus and porcine wreath virus type 2 also. IHCand IFA results also supported the single resistance to all above viruses, and the vaccination in ST cells infected with PRV showed specific reaction, which proving that these monoclonal antibody had good specificity and sensitivity. Preparatiom of PRV monoclonal antibody could provide valuable tools for further development of antigen epitope analysis and specific diagnosis reagent. |
| Key words: Pseudorabies virus monoclonal antibody specificity sensitivity |
