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Molecularly imprinted polymer test strip used for detecting 17 beta-Estradiolum and preparing method thereof

A technology of molecular imprinting and estradiol, applied in biological testing, chemical instruments and methods, analytical materials, etc., can solve problems such as complex process, difficult antibody screening, complex sample pretreatment, etc.

Active Publication Date: 2017-02-01
山东师大瑞柏生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods require complex pretreatment of samples during actual operation, which is expensive and complicated, and is not suitable for real-time on-site determination.
Rapid detection methods are mainly immunological methods, including enzyme-linked immunosorbent assays and test strip methods. This method has high selectivity due to the specific combination of antigens and antibodies, but due to the disadvantages of difficult screening, storage, and failure of antibodies, such as Chinese patent "a kit for detecting estradiol using magnetic particle chemiluminescence immunotechnology", application number: 200910091848.9, which discloses a kit for detecting estradiol using magnetic particle chemiluminescence immunotechnology

Method used

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  • Molecularly imprinted polymer test strip used for detecting 17 beta-Estradiolum and preparing method thereof
  • Molecularly imprinted polymer test strip used for detecting 17 beta-Estradiolum and preparing method thereof
  • Molecularly imprinted polymer test strip used for detecting 17 beta-Estradiolum and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Synthesis of 17β-estradiol-HRP

[0062] The 17β-estradiol used in this example was purchased from Shanghai Ruiyong Biotechnology Co., Ltd., and the HRP was purchased from sigma-aldrich Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0063] Proceed as follows:

[0064] (1) Synthesis of 17β-estradiol-3-carboxymethyl ether:

[0065] Dissolve 300mg of 17β-estradiol in 6mL of dry dimethyl sulfoxide, add 1g of potassium hydroxide powder, stir for 5 minutes, add 300mg of bromoacetic acid, continue stirring, add ice water after reacting for 2 hours; extract and recover with ethyl acetate For unreacted 17β-estradiol, the aqueous phase was acidified with 2mol / L hydrochloric acid, and a white precipitate appeared, which was filtered through a funnel. The precipitate was washed with deionized water until neutral, and vacuum-dried at 0.08MPa.

[0066] (2) Coupling of 17β-estradiol-3-carboxymethyl ether with HRP:

[0067] ①Weigh 330mg 17β-estradiol-CME, 145mg N-hydroxysuccinamide, 26...

Embodiment 2

[0070] Synthesis of 17β-estradiol-HRP

[0071] Proceed as follows:

[0072] (1) Synthesis of 17β-estradiol-3-carboxymethyl ether:

[0073] Dissolve 1g of 17β-estradiol in 10mL of dry dimethyl sulfoxide, add 5g of potassium hydroxide powder, stir for 5 minutes, add 1g of bromoacetic acid, continue stirring, add ice water after reacting for 2 hours; extract and recover with ethyl acetate For unreacted 17β-estradiol, the aqueous phase was acidified with 2mol / L hydrochloric acid, and a white precipitate appeared, which was filtered through a funnel. The precipitate was washed with deionized water until neutral, and vacuum-dried at 0.08MPa.

[0074] (2) Coupling of 17β-estradiol-3-carboxymethyl ether with HRP:

[0075] ①Weigh 1g 17β-estradiol-CME, 0.8g N-hydroxysuccinamide, 0.5g dicycloethylcarbodiimide, add 80mL dimethyl sulfoxide, and stir for 2 hours.

[0076] ② Dissolve 8 mg of HRP in 5 mL of sodium bicarbonate solution, slowly drop the solution of ① into the HRP solution, s...

Embodiment 3

[0078] Synthesis of Molecularly Imprinted Polymer Test Strips:

[0079] The paper sheet used in this example is INYC00010 Charged nylon positively charged nylon membrane NY+ purchased from Millipore Corporation (Billerica, USA).

[0080] Proceed as follows:

[0081] (1) Activation test strips

[0082] Dissolve 100mg of 3-(methacryloyloxy)propyltrimethoxysilane in 10mL ethanol solution, then soak the blank test strip in the prepared solution and incubate at room temperature for 1 hour. Rinse the solution to remove it, and then simply place the treated strips in an air stream to dry.

[0083] (2) Synthesis of molecularly imprinted polymers on test strips:

[0084](1) Polymerization solution contains 0.1 g 17β-estradiol (template molecule), 1014 μL 3-aminopropyltriethoxysilane (APTES, functional monomer), 963 μL tetraethoxysilane (TEOS, crosslinker) , 0.1mol / L acetic acid (catalyst) was dissolved in 12mL of acetonitrile solution, and the solution was ultrasonically treated fo...

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Abstract

The invention discloses a molecularly imprinted polymer test strip used for detecting 17 beta-Estradiolum and a preparing method thereof. The test strip is modified with the molecularly imprinted polymer, and the molecularly imprinted polymer bears specific binding sites of 17 beta-Estradiolum. The specific binding sites can not only carry out specific binding with enzyme marked 17 beta-Estradiolum, but also can carry out specific binding with 17 beta-Estradiolum. The detection is realized through the competitive binding between the 17 beta-Estradiolum to be detected, the enzyme marked 17 beta-Estradiolum and the biding sites. The molecularly imprinted polymer test strip used for detecting 17 beta-Estradiolum can realize the specific binding of the target template module and can magnify signal through the color reaction between enzyme and substrate. The method for detecting 17 beta-Estradiolum does not need complicated sample pretreatment, has simple operation, low cost, and good reliability, and can be used for the detection of 17 beta-Estradiolum in food or clinic.

Description

technical field [0001] The invention belongs to the technical field of molecular imprinting, and in particular relates to a molecular imprinting polymer test strip for detecting 17β-estradiol, a preparation method and a detection method thereof. Background technique [0002] Estradiol (estradiol, C 18 h 24 o 2 ) is a steroidal estrogen, there are two types of α, β, 17β-estradiol (17β-estradiol, 17β-E 2 ) is one of the most active substances among environmental estrogens. Due to the obvious carcinogenicity of 17β-estradiol, countries such as Europe and the United States have banned or strictly prohibited its use. However, 17β-estradiol has a growth-promoting effect. Abuse is common in developing countries, and there is a high probability of residues in animal foods. In addition, 17β-estradiol can be enriched in organisms through the food chain in the ecosystem, causing a long-term adverse effect and posing a serious threat to the health of human beings and natural organis...

Claims

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Application Information

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IPC IPC(8): C08G77/26C08G77/08G01N33/74G01N33/531B01J20/26B01J20/30
CPCB01J20/268C08G77/08C08G77/26G01N33/531G01N33/74G01N2600/00
Inventor 张鸿雁肖蕾刘兆臣张震
Owner 山东师大瑞柏生物科技有限公司
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