三聚氰胺单克隆抗体制备与鉴定

Preparation and identification of monoclonal antibody against melamine

中文关键词:  单克隆抗体  特性鉴定  酶联免疫吸附  胶体金免疫层析法诊断纸条法

英文关键词:monoclonal antibody  identification of characteristics  enzyme linked immunosorbent assay  colloidal gold immunochromatography diagnostic strip method

中文摘要:

      目的 制备高度特异的三聚氰胺单克隆抗体, 经酶联免疫吸附试验鉴定并制备胶体金试剂盒。方法 以牛血清白蛋白交联的三聚氰胺为抗原, 免疫BALB/c小鼠, 通过传统杂交瘤融合和筛选技术制备抗三聚氰胺单克隆抗体, 经纯化制备腹水, 采用酶联免疫吸附实验(enzyme linked immune sorbent assay, ELISA)对抗体进行效价测定、亚类测定、交叉鉴定和蛋白质免疫印迹试验(western blot)特异性鉴定后制备胶体金试剂盒。结果 筛选出1株抗三聚氰胺单抗, 制备的三聚氰胺胶体金试剂盒敏感值能达到30 μg/L。结论 获得了高度特异的且能稳定分泌三聚氰胺抗体的细胞株。

英文摘要:

      Objective To prepare highly specific melamine monoclonal antibody, colloidal gold kit was identified and prepared by enzyme-linked immunosorbent assay. Methods Using melamine crosslinked with bovine serum albumin as antigen, BALB/c mice were immunized, and anti-melamine monoclonal antibodies were prepared by traditional hybridoma fusion and screening techniques. Ascites were purified by enzyme linked immunosorbent assay (ELISA) for titer determination, subclass determination, cross identification and Western blot assay for specific identification of the antibodies to prepare colloidal gold kits. Rusults One anti-melamine monoclonal antibody was screened out, and the sensitivity value of the prepared melamine colloidal gold kit could reach 30 μg/L. Conclusion High specificity and stable secretion of melamine antibody cell lines are obtained.