抗MATSA单克隆抗体的制备与鉴定

发布时间：2018-05-15 10:34

本文选题：马立克氏病肿瘤相关表面抗原 + 单克隆抗体　；参考：《西北农林科技大学学报(自然科学版)》2009年06期

【摘要】：【目的】制备抗鸡马立克氏病肿瘤相关表面抗原(Marek's disease tumor-associated surface antigen,MATSA)的单克隆抗体(Monoclonal antibodies,McAb),为马立克氏病的相关研究及研制以MATSA单克隆抗体为载体的抗肿瘤药物提供材料。【方法】用纯化的MATSA免疫Balb/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,采用间接ELISA法筛选阳性杂交瘤细胞株,并对其染色体进行分析。将阳性杂交瘤细胞株接种Balb/c小鼠,制备McAb腹水,采用硫酸铵盐析法粗提McAb,并鉴定其所属亚类。【结果】获得了3株能稳定分泌MATSA McAb的杂交瘤细胞株B3、H6和D6,其培养上清液抗体效价为26~27,Balb/c小鼠接种细胞制得的腹水效价为210~211。染色体分析结果显示,3株杂交瘤细胞染色体数目均为52±2对。3株杂交瘤细胞株所分泌的McAb均为IgG 2b亚类,可与MSB1细胞反应。【结论】获得了MATSA的McAb。
[Abstract]:[objective] to prepare Monoclonal antibodies-McAbs, a monoclonal antibody against Marekhos disease tumor-associated surface antigenmata, in order to provide materials for the study of Marek's disease and the development of anti-tumor drugs with MATSA monoclonal antibody as carrier. Methods] Balb/c mice were immunized with purified MATSA. The spleen cells were fused with SP2/0 myeloma cells and the positive hybridoma cell lines were screened by indirect ELISA method and their chromosomes were analyzed. Balb/c mice were inoculated with positive hybridoma cell lines to prepare McAb ascites. McAbs were extracted by ammonium sulfate salting-out method and their subclasses were identified. [results] three hybridoma cell lines B _ 3H _ 6 and D _ 6 with stable secretion of MATSA McAb were obtained. The titer of antibody in the supernatant was 210211. the titer of antibody in the supernatant was 210211by inoculating cells into Balb / c mice. The results of chromosome analysis showed that the McAb secreted by the three hybridoma cell lines were all IgG 2b subclasses and could react with MSB1 cells. [conclusion] McAbs of MATSA were obtained.
【作者单位】：西北农林科技大学动物医学院;
【基金】：国家自然科学基金项目(30371067) 西北农林科技大学青年学术骨干支持计划项目(11241)
【分类号】：S858.31

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