Method for detecting sulfanilamide medicine and special enzyme-linked immunoassay reagent kit thereof
A technology of enzyme-linked immunosorbent reagents and drugs, which is applied in the preparation methods of peptides, biochemical equipment and methods, and food testing, etc., and can solve problems such as cumbersome processes, unsuitable for on-site monitoring and screening of a large number of samples, and complex instruments and equipment
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Embodiment 1
[0070] Embodiment 1, sulfamethazine antigen, sulfamethazine antibody and preparation thereof
[0071] 1. Synthesis of sulfamethazine antigen
[0072] 1. Sulfamethazine immunogen
[0073] Sulfamethazine (SM 2 ) as the sulfamethazine hapten.
[0074] 1) Take 104mg sulfamethazine (SM 2 ), add 8mL 0.25mol L -1 sulfuric acid aqueous solution, placed in a 4 ° C refrigerator to form a solution I (containing SM 2 );
[0075] 2) 200 mg of human serum albumin (HSA) was dissolved in 8 mL of Na 2 CO 3 solution (pH = 10), placed in a refrigerator at 4°C to form solution II (containing HSA);
[0076] 3) 38mg NaNO 2 , add 2mL of pure water to obtain solution III (containing NaNO 2 );
[0077] 4) Add solution III (containing NaNO 2 ) slowly added to solution I (containing SM 2 ), during this process, solution I (containing SM 2 ) placed in crushed ice for about 15min to obtain solution IV (containing SM 2 -NaNO 2 mixture);
[0078] 5) Solution IV (containing SM 2 -NaNO 2 Mi...
Embodiment 2
[0100] Embodiment 2, composition and preparation of the ELISA kit for detecting sulfamethazine
[0101] 1. The enzyme-linked immunosorbent assay kit consists of the following materials:
[0102] 1. The sulfa drug coating former shown in formula II described in embodiment 1;
[0103] 2. ELISA plate;
[0104] 3. Sulfamethazine monoclonal antibody: secreted by hybridoma cell line SAs of sulfonamide monoclonal antibody hybridoma cell line with preservation number CGMCC No.3393;
[0105] 4. Sulfamethazine standard substance: Sulfamethazine, the concentration of the standard solution is 0μg / L, 0.5μg / L, 1.5μg / L, 4.5μg / L, 13.5μg / L and 40.5μg / L L;
[0106] Sulfamethazine standard was purchased from Sigma Aldrich; CAS number: 57-68-1; diluted with sample diluent to the above concentrations;
[0107] 5. Goat anti-mouse secondary antibody labeled with horseradish peroxidase;
[0108] 6. Substrate chromogenic solution: composed of liquid A and liquid B, liquid A is an aqueous solution...
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