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| 雪卡毒素单克隆抗体的制备及鉴定 | |
| 引用本文： | 张彩霞,闫鸿鹏,郑杰,周丽华,张焜,赵肃清. 雪卡毒素单克隆抗体的制备及鉴定[J]. 免疫学杂志, 2011, 0(10): 890-893 |
| 作者姓名： | 张彩霞 闫鸿鹏 郑杰 周丽华 张焜 赵肃清 |
| 作者单位： | 广东工业大学轻工化工学院; |
| 基金项目： | 国家自然科学基金(20872020,20672023); 广东省自然科学基金(8251009001000005) |
| 摘 要： | 目的制备雪卡毒素的单克隆抗体并对其生物学特性进行鉴定。方法以雪卡毒素类似物莫能菌素为半抗原,分别采用混合酸酐法将其与载体牛血清白蛋白(BSA)偶联合成免疫抗原M-BSA,碳二亚胺法与卵清白蛋白(OVA)偶联合成包被抗原M-OVA。以M-BSA免疫Balb/c小鼠,取其脾细胞与Sp2/0骨髓瘤细胞融合制备杂交瘤。以自制雪卡毒素提取物为竞争抗原,间接竞争ELISA法筛选稳定分泌雪卡毒素抗体的特异性细胞株。小鼠体内诱生腹水,辛酸硫酸铵沉淀法进行纯化。采用抗体亚型鉴定试剂盒鉴定所得抗体的亚型,间接竞争ELISA法鉴定抗体的特异性和交叉性。结果获得3株稳定分泌抗雪卡毒素抗体的杂交瘤细胞株1D5、2G11、3G11,其Ig亚型均属于IgG1亚型。交叉反应结果表明3株单克隆细胞产生的抗体除与莫能菌素有较高的交叉反应外,与其他雪卡毒素类似物均无明显交叉反应。结论成功筛选到了分泌雪卡毒素抗体的细胞株,为雪卡毒素免疫分析方法的进一步研究奠定基础。 |
| 关 键 词： | 雪卡毒素 莫能菌素 单克隆抗体 间接竞争ELISA |
Preparation and identification of monoclonal antibodies against ciguatoxin |
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| ZHANG Caixia,YAN Hongpeng,ZHENG Jie,ZHOU Lihua,ZHANG Kun,ZHAO Suqing. Preparation and identification of monoclonal antibodies against ciguatoxin[J]. Immunological Journal, 2011, 0(10): 890-893 | |
| Authors: | ZHANG Caixia YAN Hongpeng ZHENG Jie ZHOU Lihua ZHANG Kun ZHAO Suqing |
| Affiliation: | ZHANG Caixia,YAN Hongpeng,ZHENG Jie,ZHOU Lihua,ZHANG Kun,ZHAO Suqing Department of Pharmacy Engineering,Faculty of Chemical Engineering and Light Industry,Guangdong University of Technology,Guangzhou 510006,China |
| Abstract: | This study aims to prepare monoclonal antibodies(McAbs) against ciguatoxin and to characterize their properties.Mixed anhydride and EDC methods were used for coupling monensin with BSA/OVA to prepare immunogen and coating antigen of CTXs respectively.Balb/c mice were immunized with artificial ciguatoxin antigen M-BSA,and hybridoma was generated with immunized mouse spleen cells and Sp2/0 myeloma cells.The hybridoma cell lines which secret McAbs against ciguatoxin were selected and subcloned 4 times.Conseque... |
| Keywords: | Ciguatoxin Monensin Monoclonal antibody ELISA |
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