Chlorpromazine hapten, artificial antigen, antibody as well as preparation method and application thereof
An artificial antigen, chlorpromazine technology, applied in chemical instruments and methods, animal/human protein, serum albumin, etc., can solve the problems of lack of chlorpromazine, chlorpromazine antibody specificity and insufficient recognition, and achieve The effect of high accuracy, high sensitivity and high specificity
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Embodiment 1
[0033] Example 1: Hapten Synthesis and Identification
[0034] 1. Synthesis of chlorpromazine hapten
[0035] The synthetic route of chlorpromazine hapten is attached figure 1 shown.
[0036] (1) Take by weighing 319mg (1mmol) chlorpromazine and dissolve in 5mL acetic anhydride, stir and dissolve fully in an ice bath;
[0037] (2) Take 2 mmol of freshly prepared acetyl nitrate (refer to the routine for the preparation method) and dissolve it in 1 mL of acetic anhydride and slowly drop it into the solution described in step (1), and react in an ice bath for 1 hour. Sodium oxide solution adjusted the pH value to 8, crystallized to obtain 2-chloro-10-(3-(dimethylaminopropyl))-7-nitro-5-oxo-10H-phenothiazine, the yield was 61 %;
[0038] (3) Take the above-mentioned 2-chloro-10-(3-(dimethylaminopropyl))-7-nitro-5-oxo-10H-phenothiazine and dissolve it in 20mL of ethanol, add 2 -Chloro-10-(3-(dimethylaminopropyl))-7-nitro-5-oxygen-10H-phenothiazine 10% (according to mass percen...
Embodiment 2
[0042] Example 2: Synthesis and identification of artificial antigens
[0043] 1. Synthesis and identification of chlorpromazine antigen (III)
[0044] Chlorpromazine antigen (III) glutaraldehyde synthesis:
[0045] (1) Take by weighing 35mg chlorpromazine hapten and 68mg BSA and be dissolved in 10mL phosphate buffer saline (PBS) solution together, stir at room temperature;
[0046] (2)) Add 40 μL of glutaraldehyde dropwise into the mixed solution described in step (1), and stir and react at room temperature for 3 hours after the dropwise addition;
[0047] (3) Put the solution reacted in step (2) into a dialysis bag, dialyze with PBS solution for 72 hours at 4°C, change the dialysate 6 times during the period, and obtain the target product, which is stored at -20°C after aliquoting.
[0048] In the above method, human serum protein, hemocyanin or ovalbumin can also be used. Referring to the method of this embodiment, the preparation method of chlorpromazine-coated antigen is ...
Embodiment 5
[0051] Preparation and Identification of Embodiment 5 Chlorpromazine Antigen
[0052] 2. Synthesis and identification of chlorpromazine antigen (II)
[0053] The diazotization method synthesis of chlorpromazine antigen (II):
[0054] (1) The chlorpromazine hapten that takes 35mg is dissolved in the hydrochloric acid of 1mL 1mol / L, and the hydrochloric acid adjustment solution pH value is 2 with the hydrochloric acid of 1mol / L;
[0055] (2) Take 1.1 mL of 1mol / L sodium nitrite solution and add it dropwise to the hapten solution within half an hour, and diazotize at 0-5°C for 1 hour;
[0056] (3) Weigh 68mgBSA and dissolve it in 10mL of PBS (phosphate buffer saline used routinely in the laboratory) solution, add the solution after the reaction in step (2) dropwise into the BSA-PBS solution, and use 1mol / L sodium hydroxide Adjust the pH value of the solution to 7.5, overnight at 4°C;
[0057] (4) Put the solution reacted in step (3) into a dialysis bag, dialyze with PBS soluti...
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