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Aldosterone derivatives and preparation method thereof, homogeneous enzyme immunoassay reagent for aldosterone

An aldosterone and derivative technology, which is applied in the field of aldosterone detection, can solve the problems of being unsuitable for the determination of large-scale clinical samples, high detection cost, complicated operation, etc., and achieves the effects of meeting clinical detection needs, high immunogenicity and high titer.

Active Publication Date: 2020-05-12
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

[0007] At present, there are many clinical methods for determining the content of aldosterone in serum, plasma or urine, mainly including enzyme-linked immunosorbent assay, chemiluminescence method, magnetic particle chemiluminescence method, enhanced chemiluminescence immunoassay, high performance liquid chromatography, etc. , but the existing methods have certain limitations, such as complex operation, poor stability, need to be equipped with special instruments, high detection cost, etc., are not suitable for the determination of large quantities of clinical samples
At present, there is a lack of aldosterone detection reagents with good stability, high sensitivity and strong specificity in the market, especially good quality automatic detection reagents

Method used

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  • Aldosterone derivatives and preparation method thereof, homogeneous enzyme immunoassay reagent for aldosterone
  • Aldosterone derivatives and preparation method thereof, homogeneous enzyme immunoassay reagent for aldosterone
  • Aldosterone derivatives and preparation method thereof, homogeneous enzyme immunoassay reagent for aldosterone

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Embodiment 1

[0070] Embodiment one: the preparation of aldosterone derivatives shown in formula (I) The synthetic route of aldosterone derivatives shown in formula (I) is as follows:

[0071]

[0072]

[0073] Specifically, the preparation method of aldosterone derivative shown in formula (I) comprises the following steps:

[0074] (1) Compound 4 reacts with methanesulfonyl halide (MsCl) and lithium chloride (LiCl) to generate compound 5;

[0075] (2) Compound 5 and The reaction generates compound 6;

[0076] (3) Compound 6 and Boc anhydride (Boc 2 O) react to generate compound 7;

[0077] (4) compound 7 reacts with HCl to generate compound 8;

[0078] (5) Compound 8 and SOCl 2 The reaction generates compound 9;

[0079] (6) Compound 9 and NaBH 4 The reaction generates compound 10;

[0080] (7) Compound 10 and maleimide, PPh 3 , DIAD reaction generates the aldosterone derivative shown in formula (I).

[0081] in, The synthetic route of is as follows:

[0082]

[0083...

Embodiment 2

[0118] Example 2: Synthesis of aldosterone immunogen

[0119] The aldosterone immunogen is formed by linking bovine serum albumin (Bovine Serum Albumin, BSA) with the aldosterone derivative shown in formula (I), and the specific steps of the synthetic method of the immunogen are as follows:

[0120] a. 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 7.4, and make buffer solution A;

[0121] b. Weigh 3 mg of BSA and dissolve it in 3 mL of the above buffer solution A at room temperature to make a BSA solution;

[0122] c. take by weighing the aldosterone derivative shown in 3mg formula (I), be dissolved in the above-mentioned buffer solution A of 300ul, make aldosterone derivative solution;

[0123] d. When the above-mentioned aldosterone derivative solution just became clear, it was added dropwise to the above-mentioned BSA sol...

Embodiment 3

[0125] Example 3: Preparation of anti-aldosterone specific antibody

[0126] Inoculate the experimental animals with the aldosterone immunogen prepared in Example 2. The experimental animals in this embodiment are rabbits as an example, and the antiserum is taken after booster immunization. The specific steps are as follows:

[0127] 1. Dilute the above synthesized aldosterone immunogen to 1.0 mg / ml with PBS to obtain an antigen solution, then mix 1.0 ml of the antigen solution with Freund's complete adjuvant, and inject the experimental animal rabbit.

[0128] 2. After 3 weeks, mix 1.0ml of the same antigen solution with Freund's incomplete adjuvant, inject once into the above-mentioned experimental animal rabbit, and then inject once every 4 weeks, a total of 6 injections.

[0129] 3. Take blood from the immunized experimental animal rabbit, separate and purify to obtain anti-aldosterone specific antibody with a titer of 1:30000-1:50000.

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Abstract

The invention discloses an aldosterone derivative, a preparation method thereof, an aldosterone homogeneous enzyme immunoassay reagent and preparation and detection methods thereof. The aldosterone derivative has a structure shown as a formula (I), an aldosterone immunogen with high immunogenicity and an antibody of the aldosterone immunogen are prepared from the aldosterone derivative, and the aldosterone homogeneous enzyme immunoassay reagent prepared from the antibody can be used for realizing high-flux and high-speed detection of aldosterone on a fully-automatic biochemical analyzer.

Description

technical field [0001] The invention relates to the technical field of aldosterone detection, in particular to an aldosterone derivative and a preparation method thereof, a homogeneous enzyme immunoassay reagent for aldosterone, a preparation method thereof, and a detection method. Background technique [0002] The structure of aldosterone (Aldosterone) is shown in formula (II): [0003] [0004] Aldosterone is a mineralocorticoid synthesized and secreted by the cells of the adrenal cortical zona globularis. It mainly acts on the renal distal convoluted tubule and renal cortical collecting duct, increasing the reabsorption of sodium ions and promoting the excretion of potassium ions. It also acts on the medulla The collecting duct promotes the excretion of hydrogen ions and acidifies the urine. Aldosterone is secreted through the renin-angiotensin-aldosterone system. When the volume of extracellular fluid decreases, the cells next to the glomerulus are stimulated to sec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07J43/00G01N33/53G01N33/531
CPCC07J43/003G01N33/53G01N33/531
Inventor 虞留明周洲
Owner 苏州博源医疗科技有限公司
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