Immunoassay For Three Metal Ions And Two Pesticides

Environmental pollution and food safety are the two main livelihood issues at present,among which metals and pesticides are of great concern to the society as the two main sources of pollution.Since environmental treatment takes time and ensuring the safety of food circulating in the market is the top priority at present,it is of great value to establish easy-tooperate,sensitive and low-cost detection methods.In this study,three metal ions,cadmium,aluminum and nickel,and two pesticides,boscalid and tebuconazole,were studied.Based on the principle of immunology,five corresponding monoclonal antibodies(Monoclonal antibody,m Ab)were prepared and developed for the detection of actual samples.The complete antigens of cadmium,aluminum and nickel metal ions were designed and synthesized,and the anti-Cd-ITCBE antibody 2F7,anti-Al-ITCBE antibody 3F3 and anti-NiITCBE antibody 4E3 were obtained by mouse immunization,serum detection,cell line screening and production of ascites,respectively,and the three antibodies were identified as high specificity and high affinity antibodies;the indirect competitive enzyme The IC50 values of the monoclonal antibodies against cadmium,aluminum and nickel were 0.202,2.582 and30.251 ng/m L,respectively,and the detection limits of the ic-ELISA method were 0.03,0.49 and 4.41 ng/m L,respectively,after optimizing the conditions.The results of the actual sample spiked recovery experiments showed that the established ic-ELISA method was usable;the quantitative method for the determination of cadmium in rice was established based on monoclonal antibody 2F7,and the quantitative detection value(c LOD)of the test strips was1.93 ng/g,and the detection range was 3.86-48.9 ng/g;the method for the determination of aluminum in flour and nickel in chocolate was established based on monoclonal antibodies 3F3 and 4E3,respectively.The qualitative methods for the determination of aluminum in flour and nickel in chocolate were established based on monoclonal antibodies 3F3 and 4E3,respectively,with the extinction value of 10 μg/g for aluminum and 1 μg/g for nickel in chocolate.The half antigens(Hapten)of boscalid and tebuconazole were synthesized by introducing carboxyl groups(-COOH),and the complete antigens were synthesized by carbodiimide method to obtain antibodies 4D10 and 2G5 against boscalid and tebuconazole,respectively,which were identified as highly specific and high affinity antibodies;by optimizing the concentration of antigen antibody and the p H,Na Cl content and chelator concentration of the standard diluent,an The IC50 values of antibodies 4D10 and 2G5 were 0.386 and 0.565 ng/m L,respectively,and the detection limits of the method were 0.065 and 0.123 ng/m L,respectively,and the spiked recovery experiments of the actual samples were carried out,and the recoveries and coefficients of variation indicated that the established ic-ELISA method was The recoveries and coefficients of variation indicated the usability of the established ic-ELISA method.A colloidal gold immunochromatographic test strip assay for boscalid was established,and the elimination value of the method was 10 ng/m L for boscalid in orange juice and 10 ng/g for boscalid in cucumber.