Universal artificial hapten and artificial complete antigen for capsaicins and application thereof

A technology of artificial haptens and complete antigens, which is applied in the field of immunochemistry to achieve the effects of strong specificity, high sensitivity and high affinity

Active Publication Date: 2016-05-04
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few reports on the universal artificial complete antigen for capsaicinoids at home and abroad

Method used

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  • Universal artificial hapten and artificial complete antigen for capsaicins and application thereof
  • Universal artificial hapten and artificial complete antigen for capsaicins and application thereof
  • Universal artificial hapten and artificial complete antigen for capsaicins and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The preparation of embodiment 1 capsaicinoid substance universal artificial hapten

[0029] Weigh 0.28 g of vanillin amine hydrochloride and dissolve it in 6 ml of tetrahydrofuran, add 0.15 g of triethylamine dropwise with stirring, and stir at room temperature for 30 min. Accurately weigh 0.15 g (0.0015 mol) of succinic anhydride and add to the above reaction solution, and stir overnight at room temperature. Add 3 mL of ethyl acetate to the reaction solution and stir at room temperature for 2 min. After filtration, the resulting precipitate is the hapten 4-[(4-hydroxy-3-methoxy)benzylamino]-4-carbonylcarboxylic acid (4-[ (4-hydroxy-3-methoxybenzyl)amino]-4-oxobutanoic acid), the molecular formula is C 12 h 15 NO 5 .

[0030] The results of NMR identification are: 1 HNMR(400MHz,DMSO)δ12.17(s,1H),8.87(s,1H),8.31(d,J=6.0Hz,1H),4.21(d,J=5.8Hz,2H),3.80(s, 3H), 2.52(t, J=6.9Hz, 2H), 2.42(t, J=6.7Hz, 2H). It is consistent with the theoretical value of the result, indic...

Embodiment 2

[0031] Example 2 Preparation of capsaicinoids universal artificial complete antigen-immune antigen

[0032] Weigh 20.3mg (about 0.08mmol) of the general artificial hapten of the above-mentioned capsaicinoids and 11.6mg (about 0.1mmol) of NHS in the reaction flask, add 400ulDMF to dissolve in the reaction flask, stir at room temperature for 30min, weigh 20.6mg (about 0.1mmol) 0.1 mmol) of DCC was dissolved in 100 ul of DMF, and the DCC / DMF solution was added dropwise to the above-mentioned reaction flask, stirred at room temperature for 4 h, and left standing overnight at 4°C. 8000rpm / 5min, take the supernatant active ester solution.

[0033] The supernatant active ester solution was added dropwise to 6ml of 7mg / ml BSA solution for reaction, and the reaction buffer was 0.2M phosphate buffer at pH 8.0. The reaction was carried out at room temperature for 4 h under magnetic stirring. The reaction solution was placed in a dialysis bag, and dialyzed with 0.01 M pH7.4 PBS at 4°C w...

Embodiment 3

[0034] Example 3 Capsaicinoids Universal Artificial Complete Antigen-Preparation of Coated Antigen

[0035] Dissolve 4.55 mg of the above-mentioned general artificial hapten of capsaicinoids in 200 μL of anhydrous DMF, then add 4.27 μL of tri-n-butylamine and 2.34 μL of isobutyl chloroformate in sequence, and stir at room temperature for 1 hour in the dark to obtain activated Capsaicinoid hapten solution.

[0036] The activated capsaicinoid hapten solution was added dropwise to 10ml of 4.5mg / ml OVA solution for reaction, and the reaction buffer was 0.2M phosphate buffer at pH 8.0. The reaction was carried out at room temperature for 4 h under magnetic stirring. The reaction solution was placed in a dialysis bag, and dialyzed with 0.01M PBS, pH 7.4 at 4°C with stirring, and the dialysate was changed every 4 hours for a total of 72 hours. That is, capsaicin artificial antigen-coated antigen is obtained. For the continuous scanning spectrum of UV-Vis spectrum, see figure 2 ,...

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PUM

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Abstract

The invention relates to a universal artificial hapten and artificial complete antigen for capsaicins and application thereof. The universal artificial hapten compound for capsaicins has the molecular structure disclosed as Formula I. The universal artificial complete antigen compound for capsaicins has the molecular structural formula disclosed as Formula II: Pr carrier protein. The artificial complete antigen can be immunized to obtain the capsaicin antibody with high affinity, high sensitivity and high specificity.

Description

technical field [0001] The invention relates to a general artificial hapten and an artificial complete antigen of capsaicin-like substances and applications thereof, belonging to the technical field of immunochemistry. Background technique [0002] Capsaicin compounds are the main chemical substances present in pepper fruits that make them pungent, mainly including more than 20 kinds of capsaicin, dihydrocapsaicin, homodihydrocapsaicin, nordihydrocapsaicin, homocapsaicin, etc. Known structurally similar compounds. Wherein capsaicin and dihydrocapsaicin account for about 90% of the total amount of capsaicinoids, and provide about 90% of the hot stimulation. With the improvement of the level of People's Holy Lake, the nutritional quality and medicinal value of capsaicinoids have gradually been discovered. In addition to being widely used in the food industry, capsaicinoids are also used in fields such as medicine and health care, biochemical materials, and biological weapons...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C233/22C07C231/02C07K14/765C07K14/77C07K14/795C07K16/44C07K16/06
CPCC07C231/02C07C233/22C07K14/765C07K14/77C07K14/795C07K16/065C07K16/44C07K19/00
Inventor 李培武杨青青张奇马飞张良晓张文唐晓倩丁晓霞
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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