Objective To establish an immunoassay method of wheat globulin for the rapid detection of allergen and potential protein adulteration. Methods Globulin was extracted and purified from wheat germ powder. Balb/c mice were immunized for 4 times, and the mice spleen cells and myeloma cells SP2/0 were fused as the routine cell-fushion technology. The monoclonal antibodies were obtained by immune and hybridoma technology after purification of ascites. The polyclonal antibody against wheat globulin was also prepared from rabbit serum immunized by antigen. The double antibody sandwich ELISA for wheat globulin was successfully established by optimizing parameters. Results The results showed that the titer of purified monoclonal antibody of wheat globulin was over 1: 107, and the polyclonal antibody titer was about 1:2.4×105. The minimum detection limits of enzyme linked immunosorbent assay (ELISA) kit was about 10 ng/mL, and no cross reaction was observed among proteins of different species. Conclusion The double antibody sandwich ELISA method was sensitive, specific, and provided a theoretical foundation for detection of wheat ingredients adulteration and allergens in dairy products.
标题：小麦球蛋白单克隆和多克隆抗体制备及建立酶联免疫吸附法快速检测技术
英文标题：Study on monoclonal antibody and polyclonal antibody preparation and establishment of ELISA detection method of wheat globulin
