Analgin artificial antigen synthesis method
A technology of artificial antigen and synthesis method, which is applied to the preparation method of peptides, chemical instruments and methods, animal/human protein, etc., which can solve the problems of complex pretreatment, labor and time consumption, and achieve the effect of simple synthesis steps
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Embodiment 1
[0029] Example 1 Preparation of Analgin Artificial Antigen H1-BSA
[0030] Weigh 2.03 mg of H1, dissolve it in 500 μL of ultrapure water, add 60 μL of 2.5% glutaraldehyde solution, and stir in an ice bath for 15 min to obtain the reaction solution A. Weigh 5 mg of BSA and dissolve it in 1 mL of 0.01M phosphate buffered saline solution (PBS, pH=7.4) to obtain liquid B. Add solution A to solution B dropwise, stir while adding, and react with stirring at room temperature for 2 h to obtain a coupling mixture. Take a 10 cm dialysis bag (protein cut-off: 3kDa), boil for 5 min, cool to room temperature, add the coupling mixture into the dialysis bag, and dialyze with pH=7.4, 0.01M PBS for 3 days, changing the dialysate every 6 h , to remove redundant small molecular compounds, to obtain analgin artificial antigen H1-BSA, and use the ultraviolet absorption scanning method for identification, the specific identification diagram is as follows figure 1 shown.
[0031] figure 1 Among ...
Embodiment 2
[0035] Example 2 Preparation of Metamizole Artificial Antigen H1-HS-KLH
[0036] (1) Synthesis of Sulpyrine Hapten H1-HS:
[0037]
[0038] Weigh 50 mg of 4-aminoantipyrine H1, 175 mg of 4-dimethylaminopyridine, and 300 mg of succinic anhydride, dissolve in 20 mL of anhydrous pyridine, and reflux at 60°C for 12 h. The reaction solution was concentrated by evaporation, and the residue was dissolved in 10 mL of water, extracted three times with 5 mL of ethyl acetate, and the organic phase was collected. After the organic phase was blown dry with nitrogen, a white solid was obtained, which was the analgin hapten H1-HS. Its structure was identified by NMR and mass spectrometry. The specific structure diagram is as follows: figure 2 shown.
[0039]NMR identification results: 1 H NMR (400 MHz, DMSO-d6) δ 12.09 (s, 1H, COOH), 9.08 (s,1H, NH), 7.52–7.48(m, 2H, H-1 / H-5 PhPh), 7.36–7.29 (m, 3H, H-2 / H-3 / H-4 PhPh), 3.03 (s, 3H, N-methyl), 2.53–2.46 (m, 4H, methylene), 2.09 (s, 3H,...
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