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猪繁殖与呼吸综合症病毒单克隆抗体的制备和鉴定
李建,谢红玲
0
(武汉中博生物股份有限公司)
摘要:
以猪繁殖与呼吸综合症病毒(PRRSV)作为免疫原,免疫BALB/c小鼠,融合之后经间接ELISA、IPMA和IFA试验进行杂交瘤筛选,共获得2株能分泌针对PRRSV单克隆抗体(简称单抗)的杂交瘤细胞株,分别命名为3D10和4H11 ,3D10亚类为IgG1,4H11亚类为IgG2b,单抗腹水的间接ELISA效价均达到1.0?107 ,染色体数目介于90~110之间,2株单抗与猪细小病毒(PPV)、猪伪狂犬病毒(PRV)、猪圆环病毒2型(PCV2)均无交叉反应,IPMA和IFA结果显示单抗均能与接种于猴肾细胞(Marc145)的PRRSV发生特异性反应,证实其产生的单克隆抗体具有良好的特异性和敏感性。
关键词:  猪繁殖与呼吸综合症病毒,单克隆抗体,特异性,敏感性
DOI:
投稿时间:2013-08-08修订日期:2013-11-21
基金项目:“十二五”农村领域国家科技计划课题“家畜病毒病基因工程疫苗创制”(2011AA10A208) ,廖园园,朱微,曹娟,秦伟,漆世华,谢红玲*,杨雷
Preparation and Characterization of Monoclonal Antibody against PorcineReproductive and Respiratory Syndrome Virus
(Wu Han Chopper Biological Co.,Ltd.)
Abstract:
By taking PRRSV as immunogen, BALB/c mice as model and using indirect ELISA、IPMA and IFA for hybridoma screening, we found out that 2 hybridoma cell line strains can secrete PRRSV monoclonal antibody (hereafter referred to as McAb), named 3D10 and 4H11. 3D10 subgenera is for IgG1, 4H11 subgenera for IgG2b. Single anti ascites of indirect ELISA valence achieved 1:1. 0 x 107. The chromosome number between 90 ~ 110 .These two strains showed single resistance to porcine parvovirus (PPV), porcine pseudo rabies virus (PRV) and porcine wreath virus type 2 (PCV2) also. IPMA and IFA results also supported the single resistance to all the above said viruses, and vaccination in Marc145 cells infected with PRRSV, showed specific reaction, proving that these monoclonal antibody has good specificity and sensitivity.
Key words:  porcine  reproductive and  respiratory syndrome  virus, monoclonal  antibody, specificity, sensitivity

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