| 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. |
| 关键词: 马立克氏病肿瘤相关表面抗原 单克隆抗体 杂交瘤细胞株 |
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| 基金项目: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） |
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| Preparation and identification of monoclonal antibodies against MATSA |
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| Abstract: |
| 【Objective】 The research was to obtain monoclonal antibodies against Marek′s disease tumor associated surface antigen（MATSA）to provide material for further studies on anti-tumor drugs of Marek's disease. 【Method】 Balb/c mice were immunized with purified MATSA.The spleen cells of mice were isolated and fused with SP2/0 cells.Then the positive masculine hybridoma cells were screened by indirect ELISA and its Karyotype was analysed.Finally,Balb/c mice were injected positive Hybridoma cells and the monoclonal antibody in the ascites was purified by ammonium sulfate salting-out and identified for its subtypes. 【Result】 Three hybridoma cell strains named B3,H6,D6 were obtained,and they could secrete monoclonal antibody (McAb) against MATSA steadily.McAb titers were 26-26in culture media and 210-211 in ascites.The isotyping analysis showed that these McAb all belonged to IgG 2b,and all reacted to MSB1 cell.Chromosome analysis showed that chromosome hybridoma cells numbered 52±2.【Conclusion】 McAb against MATSA was obtained. |
| Key words: MATSA monoclonal antibody hybridoma cell strain |
