抗猪圆环病毒2型Cap蛋白单克隆抗体杂交瘤细胞株的建立

Preparation of Monoclonal Antibody to the Recombinant Capsid of Porcine Circovirus Type 2

本试验旨在为PCV2的快速诊断提供技术手段。用纯化的原核表达的重组猪圆环病毒2型(PCV2)Cap蛋白(pET-Cap)免疫BALB/c小鼠,通过杂交瘤技术获得了2株能够稳定分泌抗pET-Cap抗体的杂交瘤细胞株3D5和5C5。经建立的间接ELISA试验检测3D5、5C5细胞上清效价分别为1∶2048、1∶1024;诱生腹水的效价分别为1∶204800、1∶102400,杂交瘤染色体数目分别为95条、96条。Western blotting结果表明,2株单抗能与病毒发生特异性反应。传代培养20代,3个月内复苏冻存3次,3D5、5C5能够稳定分泌抗体。

These McAbs would be useful tools for diagnostic of the PCV2 virus.BALB/c mice were immunized with the purified recombinant capsid（pET-Cap） protein of porcine circovirus type 2（PCV2） prepared from Escherichia expression system.Two monoclonal antibodies（McAbs） designated 3D5 and 5C5 were obtained.The titers of two strain McAb in cell supernatant was 1∶2048,1∶1024 and that of ascites was 1∶204800,1∶102400 respectively by indirect ELISA,the average number of chromosome were 95,96 respectively,two McAbs were confirmed to have the ability to PCV2 by Western blotting.Strain 3D5,5C5 hybridoma cell could also stably secret McAb after twenty serial passages and freezing and reviving three times.