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

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

Active Publication Date: 2019-01-11
苏州博源医疗科技有限公司
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  • 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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  • Oestrone derivative, immunogen, antibody, enzyme-labeled conjugate, detection reagent and preparation method thereof
  • Oestrone derivative, immunogen, antibody, enzyme-labeled conjugate, detection reagent and preparation method thereof
  • Oestrone derivative, immunogen, antibody, enzyme-labeled conjugate, detection reagent and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 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] The specific synthesis 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 them in 100mL of acetone, then add 7.9g (0.057mol) of K 2 CO 3 , Make a reaction mixture solution, and then heat the reaction mixture solution to reflux for 12 hours. The solution after the reaction was filtered, and the residue obtained after removing the solvent was purified by 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, 2.96g (11.3mmol) of triphenylphosphine (PPh 3 ), dissolved...

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 specific steps of the synthetic method of the estrone immunogen are as follows:

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

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

Embodiment 3

[0103] Example 3 Synthesis of 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 specific steps of the synthetic method of the 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, and dissolve them in 0.9L of deionized water, adjust the pH to 8.0, and prepare buffer solution A;

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

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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 structure of Estrone (E1) is shown in the following formula (IV): [0003] [0004] Estrone is a steroid hormone compound, a natural endogenous estrogen, which can be extracted from the pregnancy urine of pregnant women or pregnant horses. At present, estrone detection commonly used instrumental analysis methods, such as: gas chromatography, gas mass spectrometry, liquid chromatography, high performance liquid chromatography, etc. Although these methods have high sensitivity, the detection results are highly precise and accurate. However, there are disadvantages such as cumbersome sample pre-processing, long measurement time, special equipment, complicated operating procedures, long testin...

Claims

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Application Information

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Patent Type & Authority Applications(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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