摘要
用纯化的猪繁殖与呼吸系统综合征病毒(PRRSV)免疫Balb/c小鼠,应用淋巴细胞杂交瘤技术,使脾细胞与SP2/0骨髓瘤细胞融合,经间接ELISA筛选,4次有限稀释法克隆,得到1株能稳定分泌抗猪繁殖与呼吸障碍综合征病毒单克隆抗体的杂交瘤细胞株(3D6),经鉴定,该单抗为IgG2a亚类,杂交瘤细胞的平均染色体数目为99条,杂交瘤细胞培养上清及小鼠腹水单克隆抗体的ELISA效价分别为1∶512和1∶20480。这株单克隆抗体与伪狂犬病毒、猪丹毒、猪细小病毒均不发生交叉反应,显示出良好的特异性。连续培养20代后能稳定分泌抗体,此单克隆抗体的获得为PRRSV的研究及快速诊断方法的建立奠定了基础。
The Balb/c mouse were immunized for several times with the purified PRRSV,the mouse spleen cells immunized were fused with SP2/0 cells.By indirect enzyme linked immunity assay and four 4 sub-cloning,we got one monoclonal antibody hybridoma cells line（3D6）,it was the sub-type of IgG2a by identification,and the chromosome number was 99.The cells supernatant titer and ascites were 1∶512 and 1∶20480.This McAb did not occurred cross reaction with PRV,PPV and HCV.and showed excellent specificity.It could secret antibody stably for 20 generation.The McAb could be used to detect PRRSV antigen and would be useful for studying diagnostic methods of PRRSV.
出处
《中国畜牧兽医》
CAS
北大核心
2011年第4期74-77,共4页
China Animal Husbandry & Veterinary Medicine
基金
吉林省科技发展计划项目(20070209)
