摘要
为了制备特异性的抗绵羊肺腺瘤病毒(jaagsiekte sheep retrovirus,JSRV)内蒙株衣壳蛋白(CA)的单克隆抗体(McAb)杂交瘤细胞系,以原核表达CA蛋白为抗原,免疫BALB/c小鼠,经4次免疫后,取脾细胞与SP2/0骨髓瘤细胞经杂交瘤技术进行融合,同时以表达蛋白作为包被抗原进行特异性ELISA检测,共筛选到6株阳性杂交瘤细胞株。经过3次亚克隆后,最终得到了5株能稳定分泌抗体的单细胞克隆株;再利用CA真核表达蛋白以间接免疫荧光法,对此5株杂交瘤细胞进行进一步的特异性鉴定。结果显示,有3株具有特异性强荧光反应,也能检测到目的基因的表达产物。本试验获得了3株稳定分泌抗JSRV-NM株CA蛋白McAb的杂交瘤细胞系,为建立检测病原的特异性诊断方法、分析JSRV-NM株CA蛋白的功能及鉴定B细胞抗原表位等奠定基础。
In order to obtain the monoclonal antibody（McAb） against CA protein of JSRV-NM strain,BALB/c mice were immunized with purified CA fusion protein expressed in E.coli.Myeloma cells SP2/0 were fused with the splenocytes of the fore times immunized mice by hybridomas technique,and six specific antibody-producing hybridomas were screened by indirect ELISA with CA fusion protein.Five hybridomas cells of them could product McAb steadily after 3 cycles of cloning,and three McAbs of them showed positive reaction to the CA protein expressed in eukaryotic cells by indirect immunefluorescence tests and western blotting analysis.The results indicated that we had obtained three hybridomas cells which could product specific McAb against CA protein of JSRV-NM strain steadily.The McAb against CA protein of JSRV-NM strain developed would be useful as a basis of diagnosis and epitope identification.
出处
《中国畜牧兽医》
CAS
北大核心
2013年第1期28-32,共5页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金项目(31101788)
