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Estrone derivative, immunogen, antibody, enzyme-labeled conjugate, detection reagent and preparation method thereof

A technology of derivatives and immunogens, applied in the fields of enzyme-labeled conjugates, detection reagents and their preparation, immunogens, antibodies, and estrone derivatives, which can solve problems such as errors

Active Publication Date: 2020-04-17
苏州博源医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Patent application CN107652343A discloses the application of estradiol derivatives and their derivatives and kits in detecting estradiol content, but the reagent prepared from the estradiol derivatives is related to the commercially available known kits when detecting estradiol content The sex is only 0.983, there is a certain error

Method used

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  • Estrone derivative, immunogen, antibody, enzyme-labeled conjugate, detection reagent and preparation method thereof
  • Estrone derivative, immunogen, antibody, enzyme-labeled conjugate, detection reagent and preparation method thereof
  • Estrone derivative, immunogen, antibody, enzyme-labeled conjugate, detection reagent and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Embodiment 1. Synthesis of estrone derivatives

[0081] The chemical structure of estrone derivatives is shown in formula (I):

[0082]

[0083] The synthetic route and preparation steps of the above-mentioned estrone derivatives are as follows:

[0084]

[0085] Concrete synthetic steps are as follows:

[0086] (1) Synthesis of compound 3

[0087]

[0088] Weigh 5.0g (0.018mol) of compound 1 and 10g (0.048mol) of compound 2 and dissolve in 100mL of acetone, then add 7.9g (0.057mol) of K 2 CO 3 , to prepare a reaction mixture solution, and then the reaction mixture solution was heated to reflux for 12 hours. The solution after the reaction was completed was filtered, and the residue obtained after removing the solvent was purified through a silica gel purification column to obtain 3.0 g of compound 3 as a white solid.

[0089] (2) Synthesis of estrone derivatives

[0090]

[0091] Weigh 3.0g (7.54mmol) of compound 3, 1.1g (11.3mmol) of compound 4, 2.96...

Embodiment 2

[0093] Example 2. Synthesis of Estrone Immunogen

[0094] The estrone immunogen in this example is formed by linking the estrone derivative shown in the formula (I) of Example 1 with bovine serum albumin (BSA), and its structural formula is shown in the following formula (II):

[0095]

[0096] The carrier is bovine serum albumin.

[0097] The concrete steps of the synthetic method of this estrone immunogen are as follows:

[0098] (1) Weigh 2.72g of potassium dihydrogen phosphate, 4.26g of disodium hydrogen phosphate, 8.5g of sodium chloride, and 0.95g of magnesium chloride, dissolve them in 1L of deionized water, adjust the pH to 8.2, and make buffer solution A;

[0099] (2) Weigh 3mg of BSA and dissolve it in 3mL of the above buffer solution A at -4°C to make a 1mg / ml carrier solution;

[0100] (3) Weigh 3 mg of the estrone derivative represented by the above formula (I), and dissolve it in 300 μl of the above buffer solution A at -4°C to prepare a 10 mg / ml estrone der...

Embodiment 3

[0103] Synthesis of Example 3 Estrone Immunogen

[0104] The estrone immunogen in this example is formed by linking the estrone derivative shown in the formula (I) of Example 1 with bovine serum albumin (BSA), and its structural formula is shown in the following formula (II):

[0105]

[0106] The carrier is bovine serum albumin.

[0107] The concrete steps of the synthetic method of this estrone immunogen are as follows:

[0108] (1) Weigh 2.3g of potassium dihydrogen phosphate, 3.8g of disodium hydrogen phosphate, 8.0g of sodium chloride, and 0.7g of magnesium chloride, dissolve them in 0.9L of deionized water, adjust the pH to 8.0, and make buffer solution A;

[0109] (2) Weigh 6mg of BSA and dissolve it in 3mL of the above buffer solution A at -4°C to make a 2mg / ml carrier solution;

[0110] (3) Weigh 2.4 mg of the estrone derivative represented by the above formula (I), and dissolve it in 300 μl of the above buffer solution A at -4°C to prepare an 8 mg / ml estrone deriv...

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Abstract

The invention relates to an oestrone derivative for detecting oestrone content in a biological sample and a preparation method of the oestrone derivative. The structural formula of the oestrone derivative is shown as a formula (I). Oestrone immunogens with high immunogenicity and corresponding antibodies are prepared in succession by using the oestrone derivative. The oestrone immunogen is high inspecificity and high in immunogenicity, and the anti-oestrone specific antibody is high in specificity and high in titer, is capable of well recognizing and binding oestrone, and does not have any cross reaction with common 92 chaff interferents. The invention further discloses an oestrone enzyme-labeled conjugate, and an oestrone homogeneous enzyme immunoassay detection reagent is prepared by utilizing the anti-oestrone specific antibody and the oestrone enzyme-labeled conjugate. The reagent is capable of conveniently, rapidly, and accurately measuring the oestrone content in the biologicalsample, multiple samples are determined on a full-automatic biochemical analyzer, high throughput and rapid determination of the oestrone is realized, the accuracy is high, the specificity is high, and the accuracy and detection efficiency are greatly improved compared with the previous detection.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and specifically relates to an estrone derivative, an immunogen, an antibody, an enzyme-labeled conjugate, a detection reagent and a preparation method thereof. Background technique [0002] The chemical structural formula of estrone (Estrone, E1) is shown in the following formula (IV): [0003] [0004] Estrone is a steroidal hormone compound, which is a natural endogenous estrogen, which can be extracted from the urine of pregnant women or pregnant horses. At present, estrone detection methods are commonly used for instrumental analysis, such as: gas chromatography, gas chromatography-mass spectrometry, liquid chromatography, high performance liquid chromatography, etc. Although these methods have high sensitivity, the detection results have high precision and good accuracy. However, there are defects such as cumbersome sample pretreatment, long measurement time, need to be equ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07J43/00C07K14/765C07K14/77C07K14/795C07K14/435C07K16/26C07K16/06G01N33/535
CPCC07J43/003C07K14/435C07K14/765C07K14/77C07K14/795C07K16/065C07K16/26C07K19/00G01N33/535
Inventor 虞留明谢江涛陈学东郭进京
Owner 苏州博源医疗科技有限公司
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