| 三聚氰胺单克隆抗体的制备及鉴定
| Production and Identification of Monoclonal Antibodies to Melamine
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| DOI：10.3969/j.issn.1673-1689.2011.01.027
| 中文关键词: 三聚氰胺 单克隆抗体 半抗原
| 英文关键词: melamine monoclonal antibodies hapten
| 基金项目:杭州市社会发展科研攻关项目(20091133B16)
| 作者
| 单位
| 李康
| 杭州市质量技术监督检测院,浙江,杭州,310022
| 桑丽雅
| 杭州南开日新生物技术有限公司,浙江,杭州,310022
| 卜令杰
| 杭州南开日新生物技术有限公司,浙江,杭州,310022
| 陈育旺
| 杭州市质量技术监督检测院,浙江,杭州,310022
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| 中文摘要:
| 为建立三聚氰胺免疫分析方法,采用戊二醛法合成免疫抗原MEL-BSA和包被抗原MEL-OVA,全抗原经紫外吸收法检测证实偶联成功且偶联比约为16∶1。用MEL-BSA免疫BALB/c小鼠,用常规单克隆制备技术筛选获得1株稳定分泌抗三聚氰胺单克隆抗体的细胞株6E3。腹水经纯化后,研究了抗体效价、竞争抑制能力及与结构类似物的交叉反应率,其中抗体效价为1∶64 000,检测线性范围为0.1~3.2 mg/L,IC50为1.01 mg/L,与三聚氰酸、三聚氰酸一酰胺及三聚氰酸二酰胺的交叉反应率均小于0.1%,该方法有望用于三聚氰胺免疫检测试剂的研制开发。
| 英文摘要:
| In order to establish the immunoassay method of melamine(MEL),two artificial antigens(MEL-BSA,MEL-OVA) were prepared by glutaraldehyde crosslinking method,which were identified by UV absorbency spectrum.MEL-BSA was used as immuneogen to produce a cell strain(6E3) secreting monoclonal antibody against melamine from BALB/c.After purification,its titers of ascetic fluids were 64000 by indirect ELISA.The lineal range and half-inhibitory concentration(IC50) of the monoclonal antibody were from 0.1 to 3.2 mg/L and 1.01 mg/L.The percentages of cross-activity to cyanuric acid,Ammelid and Ammeline were all less than 0.1%.Results indicated that the monoclonal antibody against melamine would be useful in the development of melamine residue ELISA test kit or colloid gold-immunochromatography assay strip.
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