小鼠抗兔IgG单克隆抗体的制备与应用  

Preparation and Application of A Rat Hybridoma Cell Line Secreting Monoclonal Antibody Against Normal Rabbit IgG

作　　者：张葵[1] 汪涛[1] 李琦[1] 庄然[1] 宋朝君[1] 李永明[1] 金伯泉[1] 

机构地区：[1]第四军医大学免疫学教研室,陕西西安710032

出　　处：《标记免疫分析与临床》2008年第1期47-49,52,共4页Labeled Immunoassays and Clinical Medicine

基　　金："十一五"国家科技支撑计划重点项目;"科研用试剂核心单元物质及共性关键研制与开发"第五分题

摘　　要：制备小鼠抗兔IgG的mAb,经标记HRP和FITC后,应用于ELISA、Western blot、免疫组织化学染色试验以及FCM。以正常兔IgG为抗原,免疫BALB/c小鼠,通过常规B淋巴细胞融合技术,获得两株分泌抗兔IgG mAb的杂交瘤细胞株,间接ELISA腹水效价均达1×10-7,且亚类均为IgG1(κ)。抗体经鉴定、纯化后,以改良的过碘酸钠法标记HRP,本室常规方法标记FITC。FITC标记的2A1 mAb的F/P值为4.1,2F11的F/P值为2.7。HRP标记抗体的效价和工作浓度分别:2A1为1:25600和1:3200,2F11为1:102400和1:25600。在相同工作浓度条件下,ELISA、Western blot的实验结果,HRP标记的2F11 mAb明显优于商品化抗兔pAb,而且,2A1和2F11 mAb经标记后还可用于免疫组化和FCM,显示出很好的应用前景。Two hybridoma cell lines named 2A1 and 2F11 secreting mAb against normal rabbit IgG were established. The Ig subclass of the two mAbs was IgGl（k）. The ascites tires of the two mAbs reached to 1× 10^-7 identified by indirect ELISA. The mAbs labeled with horseradish peroxidase （HRP） or fluorescein isothiocyanate （FITC） were employed in ELISA, Western Blot, immunohistochemistry staining assay （IHC） and flow cytometry （FCM）. The F/P ratio of FITC-labeled 2A1 mAb and 2F11 was 4.1 and 2.7 respectively. The working concentrations of HRP-labeled 2A1 mAb and 2F11 were 1：3200 and 1：25600 respectively. The results of ELISA and Western blot showed that the HRP-labeled 2F11 mAb was much better than the commercial pAb. Moreover, the labeled 2A1 mAb and 2F11 could be used in IHC and FCM. The successful development of mAb against normal rabbits IgG provides a useful tool for further studying.

关 键 词：单克隆抗体 兔IGG 辣根过氧化物酶 异硫氰酸荧光素 

分 类 号：R392[医药卫生—免疫学]