胶体金标记单核细胞增生李斯特菌单克隆抗体的制备

Preparation of Monoclonal Antibodies Against Listeria Monocytogenes Coupled with Colloidal Gold

目的制备胶体金标记单核细胞增生李斯特菌的单克隆抗体(McAb),用于检测食品中的致病性单核细胞增生李斯特菌。方法以纯化的单核细胞增生李斯特菌(标准菌株号54001、54002)为抗原,免疫Balb/c小鼠,制备单克隆抗体,建立胶体金标记的单核细胞增生李斯特菌单克隆抗体的制备方法。结果筛选出2株单抗杂交瘤细胞株F5、F9,并获得免疫球蛋白,测定免疫球蛋白亚类为IgM。通过选用柠檬酸钠法制备胶体金标记单核细胞增生李斯特菌抗体,测定其最适蛋白结合浓度为12 g/ml。结论成功制备了胶体金标记的单核细胞增生李斯特菌的单克隆抗体,为建立食品中病原性单核细胞增生李斯特菌检测技术奠定基础。

Objective To prepare monoclonal antibodies against Listeria monocytogenes which was coupled with colloidal gold , to determine Listeria monocytogenes in food. Methods The Balb/c mice were immunized with the purified Listeria monocytogenes used as an antigen, monoclonal antibodies against Listeria monocytogenes were prepared by normal hybridoma technology, then the colloidal gold was coupled to the Listeria monocytogenes antibodies. Results The immuno - globulin IgM was prepared and the best compatible protein concentration of the colloidal gold was measured as 12 μg/ ml. Conclusion Listeria monocytogenes monoclonal antibodies coupled with colloidal gold have been successfully obtained, so it will build a strong basis for the detection of pathogenic Listeria monocytogenes in the future.