抗绿脓杆菌毒素A单克隆抗体杂交瘤细胞系的建立

Establishment of Hybridoma Lines Secreting Monoclonal Antibodies specific for Pseudomonas Aeruginosa Exotoxin A

本文采用绿脓杆菌外毒素A免疫BALB/c小鼠取脾细胞与Sp2/0细胞融合,建立了6株分泌单克隆抗体的杂交瘤,分别命名为14F10,23B9,23F1,23C12,,24A6及24B5。其中,23F1及24A6在传代培养中逐渐丧失了分泌抗体的能力,其余4株可稳定地分泌抗体。它们的Ig亚类鉴定;14F10为IgM,23B9为IgC1,23C12为JgG2b,24B5为JgG1。注射入同系小鼠腹腔,可产生高滴度抗体,在夹心ELISA试验中己证实均为抗绿脓杆菌外毒素A特异性的。此外,文中还讨论了在间接ELISA初次筛选中出现的假阴性及假阳性的原因及其排除方法。

From the fusion of spleen cells from the immunized mice with Sp2/0 cells were derived six hybridoma lines secreting monoclonal antibodies (McAb), designated respectively 14Flo, 23B9, 23F1, 23C12 24A6 and 2486, among which 23Fl and 24A6 lost gradually their ability to secrete antibodies during successive generations, but the rest could continously secrete McAb. Their immunoglobin isotype subclasses belong to IgM (14F10), IgG1 (23B9), IgG2b (23C12) and IgG1 (24B5). In the same inbred mice they could produce high titer antibodies which had been identified to be specific for Pseudomonas aeruginosa exotoxin A in shanwich ELISA. In addition, the causes and elimination of the false negative and positive appearing in the first scceening in indirect ELISA were discussed in the paper.