抗牛边缘无浆体MSP5单克隆抗体的制备  

Preparation of monoclonal antibodies against the MSP5 of Anaplasma marginale

作　　者：倪宏波[1,2] 姜海芳[2] 周玉龙[2] 王春仁[2] 辛九庆[3] 宫大庆[2] 钱爱东[1] 

机构地区：[1]吉林农业大学动物科技学院,吉林长春130118 [2]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [3]中国农科院哈尔滨兽医研究所,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2009年第10期819-821,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：兽医生物技术国家重点实验室开放课题基金(SKLVBF200807)

摘　　要：为制备抗牛边缘无浆体膜表面蛋白(MSP5)单克隆抗体(MAb),以原核表达的重组MSP5蛋白(rMSP5)免疫BALB/c鼠,应用常规杂交瘤技术获得2株能稳定分泌特异性MAb的杂交瘤细胞株(1D8和2F3)。间接ELISA检测腹水效价分别为5.49×105和7.83×104,亚类鉴定其重链为分别为IgG2b和lgG2a,轻链均为κ型;ELISA叠加试验表明这2株MAb识别的抗原位点相同或相近;Western blot结果显示2株MAb均能与rMSP5发生反应。特异性抗MSP5的MAb的获得,为牛边缘无浆体检测奠定了基础。Monoclonal antibodies （MAbs） against MSP5 protein of Anaplasma marginale were produced by fusing mice myeloma cell with spleencytes of BALB/c mice immunized with recombinant MSP5 protein. Two hybridoma cell lines 1D8 and 2F3 that stably produce antibodies against MSP5 were identified by ELISA. The ascites antibody titers of the hybridoma lines were 5.49×10^5 and 7.83×10^4 respectively. The sub.types of MAbs were identified as IgG2b and IgG2a. Additive index of ELISA competitive binding assay indicated that both MAbs were able to recognize identical epitopes of the MSP5 protein of A.marginale. Westem blot analysis showed that the MAbs specifically recognized certain antigenic epitopes of MSP5 protein of A.marginale, but not those of other reference antigens.

关 键 词：边缘无浆体 MSP5 单克隆抗体 

分 类 号：S852.4[农业科学—基础兽医学]