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(westernblot)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30μg/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。
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.
作者
封青
李研
孙晶莹
梁导艳
赵彭花
FENG Qing;LI Yan;SUN Jing-Ying;LIANG Dao-Yan;ZHAO Peng-Hua(Shaanxi Provincial People’s Hospital,Xi’an 710068,China;The Third Affiliated Hospital of Medical College of Xi’an Jiaotong University,Xi’an 710068,China)
出处
《食品安全质量检测学报》
CAS
2020年第15期5303-5308,共6页
Journal of Food Safety and Quality
基金
陕西省自然科学基础研究计划(2020JM-655)
陕西省科技创新支撑计划(2018-HBGC-28)
国家重点研发计划项目(2016YFD0500700)。
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
monoclonal antibody
identification of characteristics
enzyme linked immunosorbent assay
colloidal gold immunochromatography diagnostic strip method
