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采用人恶性淋巴瘤细胞株Raji细胞作为免疫原免疫BALB/C小鼠 ,按常规方法融合并经间接ELISA法筛选 ,成功地制备了 1株抗Raji细胞单克隆抗体 (MAb) ,命名为SZ1 38。用琼脂糖双扩散鉴定其为IgG1 类 ,ELISA法测定其腹水效价为 1 :3.2× 1 0 4。MAb流式细胞仪检测表明 ,该抗体与Raji和Daudi呈阳性反应 ;与外周红细胞、白细胞 (包括淋巴细胞和中性粒细胞 )、未活化血小板、内皮细胞株ECV呈阴性反应。免疫组化SP法显示 ,MAbSZ1 38识别的抗原在胃肠道的腺体和肝脏有所表达 ,而在心、脑、扁桃体、脾、脂肪组织、动脉血管壁、神经节、横纹肌、平滑肌等组织不表达或表达很低。Westernblotting分析显示 ,MAbSZ1 38可特异识别还原条件下相对分子量为 1 70kD的蛋白多肽。结果提示 :MASZ1 38对恶性淋巴瘤的诊断和治疗可能具有一定的临床意义。
BALB / C mice were immunized with human Raji cells as immunogen and fused by conventional methods and screened by indirect ELISA. One monoclonal antibody against Raji cells (MAb) was successfully prepared and named as SZ1 38 . It was identified as IgG1 by double diffusion of agarose. The ascites titer was 1: 3.2 × 10 4 by ELISA. The results of MAb flow cytometry showed that the antibody was positive to Raji and Daudi and negative to peripheral erythrocytes, leucocytes (including lymphocytes and neutrophils), non-activated platelets and ECV. The immunohistochemical SP method showed that the antigen recognized by MAbSZ1 38 was expressed in the glands and livers of the gastrointestinal tract but not in the tissues such as heart, brain, tonsil, spleen, adipose tissue, arterial wall, ganglion, striated muscle, smooth muscle Low expression or expression. Westernblotting analysis showed that MAbSZ1 38 could specifically recognize the protein polypeptide with relative molecular weight of 170 kD under reducing conditions. The results suggest that: MASZ1 38 in the diagnosis and treatment of malignant lymphoma may have some clinical significance.