抗呼吸道合胞病毒单克隆抗体的制备及其病毒蛋白特异性的识别

Development of Monoclonal Antibodies to Respiratory Syncytial Virus and Primary Identification of Their Virus Protein Specificity

应用呼吸道合胞病毒(RSV)武汉地方株滴鼻加用病毒感染的Hela细胞免疫Balb/C鼠,取脾细胞与来自实体瘤的SP2/0细胞融合,培育出分泌抗RSV单克隆抗体的杂交瘤细胞株8株其中3株有中和病毒作用。应用RSV感染Hela细胞作为RSV抗原建立了RSV的免疫转印方法,并应用免疫转印对各单抗的病毒蛋白特异性进行了初步识别:发现1株单抗识别RSV M蛋白,1株识别22 K蛋白,2株识别G蛋白,另外4株单抗的蛋白特异性尚待进一步研究确定。

The Balb/C mice were immunized with respiratory syncytial virus (RSV) infected Hela cells and the immunized spleen cells were fused with SP2/0 myeloma cells which were obtained from the SP2/0 solid tumor grown in Balb/C mice. Eight hybridoma cell lines secreting anti-RSV monoclonal antibodies(McAbs) were established. Three of them could neutralize virus. The Western-blot technique of RSV was established using RSV infected Hela cells as RSV antigen, which was used to determine the virus protein specificity of the. McAbs Western-blot analysis showed that one strain McAb recognized the RSV M protein, one strain recognized the RSV 22 K protein and two strains reacted with the RSV G protein. The virus protein specificity of the remaining 4 strains remains to be determined by further studies.