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| 三聚氰胺单克隆抗体制备与鉴定 | |
| 引用本文： | 王旻子,应永飞,龚云飞,方结红,陈宗伦,张明洲.三聚氰胺单克隆抗体制备与鉴定[J].安徽农业科学,2012(5). |
| 作者姓名： | 王旻子 应永飞 龚云飞 方结红 陈宗伦 张明洲 |
| 作者单位： | 1. 杭州迪恩科技有限公司,浙江杭州,310013 2. 浙江省畜产品质量安全检测中心,浙江杭州,310020 3. 中国计量学院生命科学学院,浙江省生物计量与检验检疫技术重点实验室,浙江杭州 310018 |
| 基金项目： | 浙江省重大科技专项项目，浙江省重点科技创新团队项目 |
| 摘 要： | 目的]制备抗三聚氰胺单克隆抗体,并对其效价、敏感性与特异性进行测定。方法]通过三聚氰胺人工抗原MEL-BSA免疫BALB/c小鼠以及细胞融合等方法,获得分泌抗三聚氰胺单克隆抗体的杂交瘤细胞株,并利用小鼠体内腹水诱生法制备抗三聚氰胺单克隆抗体;采用间接竞争ELISA法鉴定单克隆抗体的效价、敏感性与特异性。结果]经小鼠免疫、细胞融合与亚克隆后获得了2株分泌抗三聚氰胺单克隆抗体的杂交瘤细胞,分别命名为1B12和2H9;间接ELISA检测结果显示,杂交瘤细胞1B12和2H9腹水型单抗的效价分别为1∶25 600与1∶12 800,IC50分别为8.0 ng/ml与8.3 ng/ml,且该2种单抗只与三聚氰胺有反应,与三聚氰胺类似物无交叉反应。结论]该研究获得了高效价、高灵敏度的三聚氰胺特异性单克隆抗体,为后续研究提供了良好基础。 |
| 关 键 词： | 三聚氰胺 单克隆抗体 |
Preparation and Identification of Anti-Melamine Monoclonal Antibody |
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| Abstract: | Objective] To prepare the anti-melamine monoclonal antibodies and determinate their titer,sensitivity and specificity.Method] The spleen cells of BALB/c mice immunized by MEL-BSA were fused with SP2/0 myeloma cells,and hybridomas secreting monoclonal antibodies against melamine were selected and cloned.The titer,sensitivity and specificity of the monoclonal antibodies were determined by indirect ELISA method.Result] Two stable hybridoma cell lines,named 1B12 and 2H9 were established.The titer of the monoclonal antibodies secreted by 1B12 and 2H9 determined by indirect ELISA were 1∶25 600 and 1∶12 800,respectively.The monoclonal antibodies had a good sensitivity with an IC50 of 8.0 ng/ml and 8.3 ng/ml and 100% cross-reactivity to melamine,and scarcely showed cross-reactivity to others.Conclusion] The anti-melamine special monoclonal antibodies with high titer and sensitivity were obtained,which laid foundation for future research. |
| Keywords: | Melamine Monoclonal antibody |
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