Sulfamethazine antigen, preparation method and application thereof
A technology of sulfamethazine and antigen, applied in the field of sulfamethazine antigen and its preparation and application, can solve problems such as affecting the recognition of animal body, achieve improved probability, high purity and yield, and cross-reaction rate low effect
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Embodiment 1
[0022] Embodiment 1, preparation sulfamethazine hapten
[0023] 1) Dissolve 1.0g of sulfamethazine in 80mL of acetone, add 10mL of anhydrous pyridine, and mix until it is completely dissolved;
[0024] 2) Dissolve 2.0g of maleic anhydride in 20mL of anhydrous acetone, and the mixed solution obtained in 1), and reflux in a water bath at 60°C for 2 hours, the color of the solution gradually turns red, until the end of the reaction, the color of the reaction solution turns black;
[0025] 3) Evaporate acetone and pyridine to dryness to obtain a black oily substance, add 100mL 5% (mass percentage) sodium carbonate solution, and wash twice with 20mL dichloromethane;
[0026] 4) Adjust the pH to neutral, add 50 mL of ethyl acetate for extraction, discard the aqueous phase, and dry under reduced pressure to obtain 825 mg of the sulfamethazine hapten with a yield of 82.5%.
[0027] 2. Structural identification of sulfamethazine hapten
[0028] Gained sulfamethazine hapten, determine...
Embodiment 2
[0033] Embodiment 2, preparation sulfamethazine antigen
[0034] 1) Preparation of sulfamethazine antigen
[0035] Weigh 10 mg of the sulfamethazine hapten of Example 1 and dissolve it in 5 mL of N, N-dimethylformamide (DMF), ultrasonically dissolve it completely, and cool with cold ethanol; then add 10 mg of N, N '-Dicyclohexylcarbodiimide (DCC) and 5mg N-hydroxysuccinimide (NHS), react overnight at room temperature; finally add bovine serum albumin or ovalbumin carbonate solution dropwise (50mg bovine serum albumin or ovalbumin dissolved in 5 mL of 0.01M carbonic acid solution), and continued to stir for 4 hours; then put the reaction solution into a dialysis bag, and dialyzed with normal saline solution at 4°C for 48 hours, changing the water 6 times. The dialysate was passed through a 0.2 μm filter membrane under sterile conditions to obtain the sulfamethazine antigen (SM 2 -BSA) or coating antigen (SM 2 -OVA), aliquoted into ampoule bottles and stored at -20°C.
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