Conjugate for gentiopicroside and preparation method thereof
A technology of gentiopicroside and conjugates, which is applied in the field of gentiopicroside conjugates and its preparation, and can solve problems such as not being able to meet detection needs
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Embodiment 1
[0019] (1) Preparation of solution A: in a 50 mL round reaction flask, while stirring (120 rpm), add 17.82 mg (0.050 mmol) of gentiopicroside, 16.2 mg of N,N'-carbonyldiimidazole (CDI) And 0.6 mg of 4-dimethylaminopyridine (DMAP) was dissolved in 2 mL of dimethylformamide (DMF) and reacted for 4 hours to generate the active intermediate of gentiopicroside and CDI.
[0020] (2) Preparation of solution B: 171 mg (0.0025 mmol) of cBSA was dissolved in 15 mL of PBS (pH=7.4, 0.01 M) at 25°C. Under magnetic stirring (120 rpm), solution A was added dropwise to solution B, and the solution B was obtained by reacting for 6 hours.
[0021] (3) Preparation of gentiopicroside immunogen: the solution B was dialyzed against PBS (pH=7.40, 0.01M), and the dialysate was replaced every 8 hours for 3 consecutive days. Then, it was permeated with distilled water for 3 days, and the dialysate was changed every 8 hours. The dialysate was collected and lyophilized to obtain 150 mg of white solid p...
Embodiment 2
[0023] (1) Preparation of solution A: in a 50 mL round reaction flask, while stirring (120 rpm), add 17.82 mg (0.050 mmol) of gentiopicroside, 16.2 mg of N,N'-carbonyldiimidazole (CDI) And 0.6 mg of 4-dimethylaminopyridine (DMAP) was dissolved in 2 mL of dimethylformamide (DMF), and reacted for 6 hours to generate the active intermediate of gentiopicroside and CDI.
[0024] (2) Preparation of solution B: 25.72 mg (0.0033 mmol) of cBSA was dissolved in 15 mL of PBS (pH=7.4, 0.01 M) at 28°C. Under magnetic stirring (120 rpm), solution A was added dropwise to solution B, and the solution B was obtained by reacting for 6 hours.
[0025] (3) Preparation of gentiopicroside immunogen: the solution B was dialyzed against PBS (pH=7.40, 0.01M), and the dialysate was replaced every 8 hours for 3 consecutive days. Then, it was permeated with distilled water for 3 days, and the dialysate was changed every 8 hours. The dialysate was collected and lyophilized to obtain 165 mg of white soli...
Embodiment 3
[0027] (1) Preparation of solution A: in a 50 mL round reaction flask, while stirring (120 rpm), add 17.82 mg (0.050 mmol) of gentiopicroside, 16.2 mg of N,N'-carbonyldiimidazole (CDI) And 0.6 mg of 4-dimethylaminopyridine (DMAP) was dissolved in 2 mL of dimethylformamide (DMF), and reacted for 8 hours to generate the active intermediate of gentiopicroside and CDI.
[0028] (2) Preparation of solution B: 114.00 mg (0.0017 mmol) of cBSA was dissolved in 15 mL of PBS (pH=7.4, 0.01 M) at 30°C. Under magnetic stirring (120 rpm), solution A was added dropwise to solution B, and the solution B was obtained by reacting for 6 hours.
[0029] (3) Preparation of gentiopicroside immunogen: the solution B was dialyzed against PBS (pH=7.40, 0.01M), and the dialysate was replaced every 8 hours for 3 consecutive days. Then, it was permeated with distilled water for 3 days, and the dialysate was changed every 8 hours. The dialysate was collected and lyophilized to obtain 135 mg of white sol...
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