Sorbic acid hapten, artificial antigen, antibody and its synthesis method and application

A synthetic method and artificial antigen technology, applied in the field of immunochemical analysis, to achieve the effects of easy control of reaction conditions, high purity and yield, and cost advantages in synthesis

Active Publication Date: 2021-09-07
SHENZHEN BIOEASY BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, at present, there is no relevant report on sorbic acid hapten in China

Method used

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  • Sorbic acid hapten, artificial antigen, antibody and its synthesis method and application
  • Sorbic acid hapten, artificial antigen, antibody and its synthesis method and application
  • Sorbic acid hapten, artificial antigen, antibody and its synthesis method and application

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Experimental program
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preparation example Construction

[0035]Specifically, the formula ( ) The synthetic method of the sorbic acid hapten of the shown structure comprises the following steps:

[0036] S1. Under the catalysis of dipolyrhodium acetate, furan reacts with ethyl azidoacetate to obtain intermediate 1, which has the structural formula ;

[0037] S2. The intermediate 1 is subjected to a hydrolysis reaction to obtain the formula ( ) of the sorbic acid hapten with the structure shown.

[0038] Wherein, the step S1 includes the following steps: firstly add dimeric rhodium acetate and furan into the reactor, then slowly add ethyl azidoacetate dropwise under ice bath, after the dropwise addition, continue to stir the reaction at room temperature, after the reaction ends , add water, extract with ethyl acetate, collect the organic phase, evaporate the organic phase to dryness and redissolve with anhydrous dichloromethane, then add iodine particles, ultrasonically dissolve, and continue to stir the reaction at room tempera...

Embodiment 1

[0064] A kind of hapten for sorbic acid immunization, its synthesis method comprises the steps:

[0065]S1. Add rhodium acetate and furan to the reactor first, and then slowly add ethyl azidoacetate dropwise under ice bath. Carry out extraction, collect the organic phase, redissolve the organic phase with anhydrous dichloromethane after evaporating to dryness, then add iodine particles, ultrasonically dissolve, continue to stir the reaction at room temperature, add an oxidant after the reaction is completed, and evaporate the separated organic phase to dryness , column purification, to obtain intermediate 1; wherein, the ratio of the amount of the furan and ethyl azidoacetate is 2:1; the quality of the dipoly rhodium acetate is the quality of the ethyl azidoacetate 0.08%; The amount of the substance of the iodine particle is 2.5 times of the amount of the ethyl azidoacetate substance;

[0066] S2. Weigh intermediate 1, dissolve it with methanol, add alkaline solution, heat to...

Embodiment 2

[0068] A kind of hapten for sorbic acid immunization, its synthesis method comprises the steps:

[0069] S1. Add rhodium acetate and furan to the reactor first, and then slowly add ethyl azidoacetate dropwise under ice bath. Carry out extraction, collect the organic phase, redissolve the organic phase with anhydrous dichloromethane after evaporating to dryness, then add iodine particles, ultrasonically dissolve, continue to stir the reaction at room temperature, add an oxidant after the reaction is completed, and evaporate the separated organic phase to dryness , column purification, to obtain intermediate 1; wherein, the ratio of the amount of the furan and ethyl azidoacetate is 3:1; the quality of the dipoly rhodium acetate is the quality of the ethyl azidoacetate 0.1; The amount of substance of described iodine grain is 2 times of the amount of ethyl azidoacetate substance;

[0070] S2. Weigh intermediate 1, dissolve it with methanol, add alkaline solution, heat to reflux,...

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Abstract

The present invention discloses sorbic acid hapten, artificial antigen, antibody and its synthesis method and application. In the present invention, two kinds of sorbic acid haptens with different structures are provided, wherein the sorbic acid hapten with the structure shown in formula () is used as The hapten for sorbic acid immunization, the hapten for sorbic acid immunization is highly similar to the target analyte sorbic acid, retains the characteristic structure of sorbic acid to the greatest extent, and can better expose the epitope of the hapten, making sorbic acid The immunogenicity of the hapten is obviously enhanced, and it has an aldehyde group that can be coupled with the carrier protein; the sorbic acid immunization antigen obtained after the hapten is coupled with the carrier protein is used to immunize animals, which is more conducive to stimulating animals. The immune response produces antibodies with stronger specificity and higher sensitivity, and the IC of sorbic acid antibody is detected by indirect competitive ELISA 50 It is 11mg / L, which provides the basis for the subsequent establishment of various immunoassay methods for sorbic acid.

Description

technical field [0001] The invention relates to the technical field of immunochemical analysis, in particular to sorbic acid hapten, artificial antigen, antibody and their synthesis method and application. Background technique [0002] Sorbic acid and potassium sorbate are good food preservatives, which are widely used in agricultural products, food and other industries. Through the application of potassium salt and sorbic acid, the growth of microorganisms can be effectively controlled and the shelf life can be extended. In the field of food, the daily allowable amount of potassium sorbate and sorbic acid is strictly defined. Excessive consumption of these two substances will cause harm to the human body, which is not conducive to the maintenance of the balance of normal intestinal flora and is not conducive to the Normal maintenance of metabolism. In severe cases, it will lead to the inhibition of bone growth and threaten the health of the liver and kidneys. my country's...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/00
CPCC07C51/09C07C51/367C07C59/52C07C59/74C07C67/343C07K14/765C07K16/44C07K19/00G01N33/577C07C69/738
Inventor 付辉李细清杨星星王伟维杨林林曾文涛周巧妮曾楚怡张克俊艾泉
Owner SHENZHEN BIOEASY BIOTECHNOLOGY CO LTD
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