Hypoletin hapten, artificial antigen, preparation method thereof, antibody and application thereof

An artificial antigen, scopoletin technology, applied in the field of antibody, scopoletin hapten, artificial antigen and its preparation method, can solve the problems of low detection efficiency, high detection cost, etc., and achieve the effect of high titer

Active Publication Date: 2021-10-29
TOBACCO RES INST CHIN AGRI SCI ACAD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the embodiment of the present invention is to provide a scopoletin hapten, which aims to solve the problems of low detection efficiency and high detection cost in the existing scopoletin detection method question

Method used

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  • Hypoletin hapten, artificial antigen, preparation method thereof, antibody and application thereof
  • Hypoletin hapten, artificial antigen, preparation method thereof, antibody and application thereof
  • Hypoletin hapten, artificial antigen, preparation method thereof, antibody and application thereof

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preparation example Construction

[0027] The embodiment of the present invention also provides a preparation method of the above-mentioned scopoletin hapten, comprising:

[0028] Dissolving scopoletin in pyridine, adding dianhydride and dimethylaminopyridine to carry out reflux and stirring reaction, after the reaction is completed, carry out vacuum evaporation of the pyridine, dissolved residue and washing liquid washing treatment, and dry the obtained upper organic phase, reduce After concentrated under pressure, chromatographic purification, gradient eluent and concentrated treatment, it is obtained.

[0029] Wherein, the molar mass ratio of scopoletin to dianhydride is 1:2-5.

[0030] Wherein, the dianhydride is one of succinic anhydride, glutaric anhydride and adipic anhydride; preferably succinic anhydride.

[0031] Wherein, the reaction condition is stirring and heating under reflux for 6-12 hours.

[0032] Wherein, the washing liquid includes hydrochloric acid, ultrapure water, saturated saline; pref...

Embodiment 1

[0045] The present embodiment provides a scopoletin hapten, the structural formula of which is as follows:

[0046]

[0047] It can be seen that the hapten provided by the present invention directly introduces carboxyl groups through the hydroxyl groups on scopoletin, which retains the chemical structure and characteristics of scopoletin to the greatest extent, and can be directly coupled with carrier proteins, which is useful for the subsequent preparation of high The specific and highly sensitive scopoletin antibody laid the foundation.

Embodiment 2

[0049] The present embodiment provides a kind of preparation method of scopoletin hapten, and its steps are as follows:

[0050] Dissolve 0.58g (3mmol) scopoletin in 10mL anhydrous pyridine, add 1.5g (15mmol) succinic anhydride and 0.05g dimethylaminopyridine (DMAP), and react under reflux for 12h. Evaporate pyridine under reduced pressure, dissolve the residue with 10 mL of ethyl acetate, then wash with hydrochloric acid (1M, 20 mL, 3 times), ultrapure water (20 mL, 2 times), and saturated brine (20 mL, 2 times) respectively . Then the upper organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Purified by silica gel column chromatography, and carried out gradient elution with ethyl acetate:petroleum ether (volume ratio 1:1), the eluate was collected and concentrated to obtain scopoletin hapten (yield 90%). The corresponding reaction equation is as follows:

[0051]

[0052] The preparation method of the scopoletin hapten of the ...

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Abstract

The invention is suitable for the technical field of quality detection, and provides a scopoletin hapten, a scopoletin artificial antigen, a preparation method thereof, an antibody and an application thereof. The structural formula of the scopoletin hapten is shown in the specification, and n is any integer from 2 to 4. A carboxyl group is directly introduced into a hydroxyl group of the scopoletin, the chemical structure and characteristics of the scopoletin are reserved to the maximum extent, the scopoletin can be directly coupled with carrier protein, and a foundation is laid for subsequent preparation of antibodies with high specificity and high sensitivity; moreover, the scopoletin artificial antigen is prepared on the basis of the scopoletin hapten, and an antibody obtained from the scopoletin artificial antigen has high titer, specificity and affinity and can be applied to rapid and specific detection of scopoletin in agricultural products such as tobacco leaves and related food.

Description

technical field [0001] The invention belongs to the technical field of quality detection, and in particular relates to a scopoletin hapten, an artificial antigen, a preparation method, an antibody and an application thereof. Background technique [0002] Scopoletin is a coumarin derivative in plants, which is commonly found in flue-cured tobacco leaves, Burley tobacco leaves, and smoke, and also in some Chinese herbal medicines and fruits such as wolfberry, Digupi, Dawanhua, and Noni fruit. As the main component of polyphenols in tobacco, scopoletin plays an important role in the growth and development of tobacco, modulation characteristics, tobacco leaf color, aroma and taste, and is an important indicator for measuring tobacco quality. In addition, scopoletin has anti-oxidation and anti-fungal effects, and can effectively inhibit the growth of Fusarium half-naked bacteria, so it is also an important active substance in Chinese herbal medicine and food. The content of scop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D311/16C07K14/765C07K14/77C07K16/44G01N33/53
CPCC07D311/16C07K14/765C07K14/77C07K16/44G01N33/5308
Inventor 方松孔凡玉华修德刘雪宁扬宋德安邱军
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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