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恶性淋巴瘤单克隆抗体的制备和初步鉴定
Preparation and Characterization of Monoclonal Antibody Against Human Malignant Lymphoma
【摘要】 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。
【Abstract】 BALB/C mice was immunized with Raji cell suspension.The cells were fused as a normal method and the MAb was screened by indirect ELISA. A murine MAb SZ138 was obtained and its titer in ascites was 1:3.2×10 4. MAb was typed by reaction with subclass-specific rabbit anti-mouse antibody and was found to be IgG 1. Flow cytometry showed that MAb SZ138 was postive with Raji and Daudi cells, while did not react with red blood cells, white blood cells, inactive platets and ECV304 cell.Immunohistochemical SP staining confirmed that the antigen identified by MAb SZ138 was expressed on the gastroenteric duct and liver, but very little or not on cardiac, brain, tonsile, spleen etc. Western blotting demonstrated that MAb reacted with the antigen whose molecular weight was about 170kD under reducing condition.As a result, MAb SZ138 with high immuno-reactivity and tissue specificity may perhaps be useful for diagnosis and treatment of malignant lymphoma.
- 【文献出处】 标记免疫分析与临床 ,Labeled Immunoassays and Clinical Medicine , 编辑部邮箱 ,2004年01期
- 【分类号】R733.1
- 【被引频次】1
- 【下载频次】47