抗辣根过氧化物酶单克隆抗体的研制和鉴定

Development and Characterization of Mouse Antiperoxidase Monoclonal Antibody

用国产辣根过氧化物酶(HRP)免疫BALB/C鼠,取免疫脾细胞与SP2/0小鼠骨髓瘤细胞融合,获得了7株分泌HRP特异性抗体的杂交瘤细胞株,其培养上清液的效价为256～4096,腹水效价为1024×10~3～4 096×10~3.上述7株细胞株中的6株所产生的单克隆抗体均能制备出质量良好的鼠单克隆PAP复合物,用于免疫组织化学及血清学研究均取得较理想效果。将7株分泌HRP特异性抗体的杂交瘤细胞株适应于含8-氮杂鸟嘌呤培养基中,诱导出了对HAT敏感的突变细胞株,为研制双特异性单克隆抗体奠定了基础。

Mouse-mouse hybridoma technique has been established successfully for development of mouse antiperoxidase monoclonal antibody. Seven hybridomas secreting monoclonal antibody against HRP were obtained. The antibody titration of supernatants was 256-4096, while that of ascites was 1 024×103-4 096× 10s. The PAP complexes prepared from these monoclonal antibodies were used in immunohistochemical studies. Also described is the production of HAT sensitive variant cell lines offering the possibility of using these hybridomas as parent cell line for the production of hybrid-hybridomas secreting bi-specific antibodies.