狂犬病病毒糖蛋白单克隆抗体的制备

摘要

用狂犬病病毒BD06株脑毒免疫BALB/c小鼠,制备G蛋白单克隆抗体.将灭活的BD06脑毒免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞(SP2/0)进行融合,通过间接ELISA法和荧光抗体病毒中和试验法进行4轮筛选,获得6F12、1B12共2株具有中和活性的糖蛋白单克隆抗体的杂交瘤细胞.通过Western blotting分析和直接免疫荧光检测结果显示,获得一株针对狂犬病病毒糖蛋白构象表位的单抗6F12和一株针对糖蛋白线性表位的单抗1B12.小鼠中和试验结果显示,两株杂交瘤细胞分泌的抗体均具有狂犬病病毒中和活性.结果表明,获得了两株针对狂犬病病毒G蛋白不同抗原表位的中和活性单克隆抗体,为狂犬病病毒特性研究和检测奠定了基础.%Two monoclonal antibodies against rabies virus glycoprotein were produced by immunization with rabies virus strain BD06 in BALB/c mice. Splenocytes from the immunized mice were fused with SP2/0 mouse myeloma cells. After fusion, antibodies in the supernatant of the hybrid cells were respectively detected by ELISA and FAVN tests. Two hybrid cells secreting antibody binding to rabies virus, i. e. 6F12 and 1B12 strains, were analyzed by Western blotting and FAT methods. Results showed that conformational and linear epitopes of the glycoprotein were repectively recognized by neutralizing monoclonal antibody 6F12 and 1B12.