小鼠杂交瘤细胞系的建立及抗兔IgG单克隆抗体的应用  

ESTABLISHMENT OF HYBRIDOMA CELL LINES SECRETING MONOCLONAL ANTYBODIES(MAB)AGAINST RABBIT IgG AND ITS APPLICATION

作　　者：王远程[1] 郭军[1] 蔡少华[1] 

机构地区：[1]中国农业科学院生物技术研究中心

出　　处：《微生物学报》1991年第5期357-363,共7页Acta Microbiologica Sinica

摘　　要：经三次免疫一次细胞融合,得到分泌抗兔 IgG 的杂交瘤细胞克隆27个。细胞融合率100%,阳性孔率4%。经筛选和克隆化培养建立了杂交瘤细胞系3C8E4和6A3H6,其免疫球蛋白分别为 IgG2a 和IgG3。3C8E4腹水抗体滴度超过1:10240000 3C8E4除和兔 IgG 有反应外,还和人与马 IgG 有反应;6A3H6只和兔 IgG 有反应,与其它七种动物和人 IgG 无交叉反应。经用过碘酸盐法将3C8E4 MAb 和辣根过氧化物酶标记,制得酶标抗体,当使用浓度为1:10000时,其OD>2.0。用该酶标记抗体 ELISA 法检测植物病毒 PVX、PVY、TEV 和TuMV 效果良好。BALB/c mouse was immunized three ti-mes and cell fusion was performed betweensplenocytes of the mice and Sp2/O-Ag-14myeloma cells after 72 hours of last immuni-zation.Fusion rate was 100% and specificpositive rate of rabbit IgG was 4%,twenty-seven specific positive clones of rabbit IgGwere obtained.Cell lines 3C8E4 and 6A3H6were established in four cloning hybridomas.The immunoglobulins of MAb-3C8E4 andMAb-6A3H6 are IgG2a and IgG3 respecti-rely.Ascites titer of 3C8E4 was over 1:1024000.Besides rabbit IgG,3C8E4 hascross reactions with human IgG and horseIgG in eight animals and human,6A3H6can only react with rabbit IgG.MAb-3C8E4was conjugated with horseradish peroxidase(HRP) by using improved sodium periodatemethod.The conjugate titer was over 1∶40000,the O.D.value was over 2.0 when thedilution was 1∶10000.It gaves a good resultwhes using ELISA methol to detect plantviruses PVX,PVY,TEV,TuMV by using theconjugate.

关 键 词：杂交瘤 单克隆抗体 免IgG 

分 类 号：Q939.91[生物学—微生物学]