猪繁殖与呼吸道综合征病毒(PRRSV)单克隆抗体的研制

Development of the Monoclonal Antibodies against Porcine Reproductive and Respiratory Syndrome Virus(PRRSV)

利用猪繁殖与呼吸道综合征病毒 ( PRRSV)国内分离株 J1 ,采用反复差速离心法制备免疫抗原 ,长程免疫法免疫 BALB/ c小鼠 ,用间接 ELISA方法检测抗体 ,通过细胞融合技术 ,并经 3次亚克隆获得了 1 0株能稳定分泌抗 PRRSV单抗的杂交瘤细胞克隆株 ( A1 D7H1 0 ,A1 D7H1 1 ,A1 E7H9,A1 E7D9,A2 D8E7,A2 D8B1 1 ,B3D1 1 D6 ,B3D1 1 E6 ,B2 G9A9,B2 G9F2 )。这些细胞经体外连续传代 1个月和冻存 2个月复苏后 ,均能稳定分泌高效价的抗 PRRSV抗体 ,并能在小鼠体内形成肿瘤 ,有效诱导产生高效价 PRRSV抗体腹水 ( ELISA效价为1 0 - 3～ 1 0 - 4 )。其中有 7株能与美洲弱毒株

Indirect enzyme linked immunosorbent assay(ELISA) method was developed to detect antibodies against PRRSV with J 1 isolated from the Huabei region of China. The viral antigens were purified by a number of centrifugations at low speed and ultraspeed. After the BALB/c mice to be immunized five times,the secreting antibody hybridomas were obtained.By screening with ELISA and cloning three times using the method of limiting dilution,ten hybridoma cell lines with a high and stable level of antibodies secreted were developed. The monoclonal antibodies were produced in cell culture supernatants or in ascites. The ELISA titer of the ascites was 10 -3 10 -4 .Among them,seven hybridoma cell lines of monoclonal antibodies of PRRSV J 1 strain could be bound to Rerp PRRSV strain.