Estradiol hapten, antigen and antibody as well as preparation methods and applications of estradiol hapten, antigen and antibody

A technology of estradiol and hapten, applied in the fields of estradiol hapten, antibody and its preparation, and antigen, which can solve problems such as competitive interference, antibody difficulty, immune tolerance and endocrine system interference, and achieve high sensitivity Effect

Inactive Publication Date: 2014-09-24
高海岗
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since estradiol is a subtype compound, it is very likely that there may be carrier determinant competitive interference, immune tolerance and endocrine system interference, and it is often difficult to prepare antibodies against it

Method used

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  • Estradiol hapten, antigen and antibody as well as preparation methods and applications of estradiol hapten, antigen and antibody
  • Estradiol hapten, antigen and antibody as well as preparation methods and applications of estradiol hapten, antigen and antibody
  • Estradiol hapten, antigen and antibody as well as preparation methods and applications of estradiol hapten, antigen and antibody

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] In this paper, DMF refers to N,N-dimethylformamide; DCC refers to N,N-dicyclohexylcarbonimide; NHS refers to N-hydroxysuccinimide; PBS refers to phosphate; KLH refers to keyhole limpet hemocyanin ; OVA refers to chicken ovalbumin; H represents estradiol hapten; H-KLH refers to estradiol antigen whose carrier protein is keyhole limpet hemocyanin; H-OVA refers to estradiol antigen whose carrier protein is ovalbumin.

[0034] Preparation method of hapten

[0035] Dissolve 10 mmol of estradiol in dichloromethane solution and add dropwise to a dichloromethane solution containing 5 mmol of glutaric anhydride, add 1 mmol of catalyst 4-dimethylaminopyridine to carry out catalytic reaction, and stir at 37°C for 1 h in the dark. After the reaction was completed, the reaction solvent was spin-dried under vacuum at 40°C, 5 mL of water was added to dissolve the unreacted estradiol and glutaric anhydride, the reaction product was extracted with 5 mL of ether, and the ether was spin-d...

Embodiment 2

[0051] Preparation method of hapten

[0052] Dissolve 10 mmol of estradiol in dichloromethane solution and add dropwise to a dichloromethane solution containing 5 mmol of glutaric anhydride, add 1 mmol of catalyst 4-dimethylaminopyridine to carry out catalytic reaction, and stir at 37°C for 1 h in the dark. After the reaction was completed, the reaction solvent was spin-dried under vacuum at 40°C, 5 mL of water was added to dissolve the unreacted estradiol and glutaric anhydride, the reaction product was extracted with 5 mL of ether, and the ether was spin-dried to obtain a white solid, which was the estradiol hapten. The rate is 86%, and the reaction formula is as follows:

[0053]

[0054] Preparation method of antigen

[0055] Active ester method: Dissolve 0.1 mmol of the hapten prepared above in 0.5 mL of DMF, add 0.2 mmol of DCC and 0.2 mmol of LNHS with stirring, and magnetically stir the reaction at 4°C for 12 hours overnight. After centrifugation, the supernatant b...

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Abstract

The invention discloses an estradiol hapten with a structural formula as shown in the specification, an antigen and an antibody as well as preparation methods and applications of the estradiol hapten, the antigen and the antibody. Most instrument means are adopted in the traditional estradiol detecting technology, and the instrument means are complex in operation and pretreatment and high in cost. The invention provides the estradiol hapten, the antigen and the antibody as well as the preparation methods and applications of the estradiol hapten, the antigen and the antibody. The hapten is prepared by extending the arm of a feature group of estradiol, and furthermore, the antigen is prepared, so that an immune animal obtains a specific antibody; the antibody can be used for specifically identifying estradiol; due to the extension of the arm of the feature group of the hapten, the distance between the hapten and a carrier protein is increased, the steric hindrance is reduced, and the immune response of animals can be favorably stimulated to generate the antibody with stronger specificity and higher sensitivity. The inhibition ratio of the specific estradiol antibody provided by the invention to estradiol is up to 91%.

Description

technical field [0001] The present invention relates to the preparation method of hapten, antigen and antibody, especially relates to estradiol hapten, antigen, antibody and preparation method and application thereof. Background technique [0002] Estradiol (17β-Estradiol) is an important component of natural estrogen, because it can significantly promote animal growth and improve the lean meat ratio of livestock and poultry, and is used in the production of livestock and poultry industry. Comparison of the half-life of estradiol in animals It can accumulate in the liver, muscle and kidney of livestock and poultry, enter the human body through the food chain, interfere with the normal endocrine system of the human body, and cause problems such as precocious puberty in children and feminization of breast development in male children. Many countries have stipulated the maximum residue limit of estradiol and its derivatives. For example, the Food Safety and Inspection Service o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07J1/00C07K14/795C07K14/765C07K14/77C07K16/44G01N33/53
Inventor 高海岗陆建荣顾菲菲张月峰马光焰施妮莎许国平
Owner 高海岗
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