抗伪狂犬病病毒gG蛋白单克隆抗体的制备及鉴定

Establishment and Characterization of Monoclonal Antibody to PRV gG protein

以大肠杆菌表达的猪伪狂犬病病毒 (PRV) g G蛋白作为抗原 ,免疫 BAL B/c小鼠 ,经细胞融合、克隆后 ,获得了1F6、2 B92株稳定分泌抗伪狂犬病病毒 g G蛋白单克隆抗体的杂交瘤细胞。 1F6、2 B9培养上清及小鼠腹水效价(EL ISA)分别为 1∶ 2 1 1 、1∶ 2 1 1 及 1∶ 10 0× 2 1 1 、1∶ 10 0× 2 1 2 。 1F6、2 B9的染色体众数分别为 87(80～ 94 )、84 (81～ 87)。经间接 EL ISA鉴定 ,1F6、2 B9单抗分别属于 Ig G2 a、Ig G3亚类。1F6、2 B9与猪伪狂犬病病毒 Ea株感染细胞的间接免疫荧光试验结果表明 ,1F6、2 B9确实为伪狂犬病病毒 g G蛋白的单抗。所得单抗与牛传染性鼻气管炎病毒 (IBRV)、猪细小病毒 (PPV)、猪繁殖与呼吸道综合征病毒 (PRRSV)

Two hybridoma lines producing monoclonal antibody against PRV gG glycoprotein have been established.After immunization of BALB/c mice with gG protein expressed in E.coli,the stimulated spleenocytes were fused with SP2/0 myelomas to produce hybridomas.Two hybridoma lines,designated 1F6 and 2B9,were isolated and expressed antibodies of subclasses IgG2a and IgG3 respectively.The number of chromosome of the two hybridoma lines was 80-94 and 81-87 respectively.The ELISA titers of cell supernatant and ascites of 1F6 and 2B9 were 1∶2^(11),1∶2^(11) and 1∶100×2^(11),1∶100×2^(11) respectively.By indirect immunofluorescence assay,the two McAb reacted with cells infected by PRV positively and did not react with cells infected by IBRV,PPV,PRRSV.The AI results of additivity ELISA indicated that the two McAbs were against the same epitope.