3-(3-hydroxyphenyl)-3-hydroxypropionic acid derivative, homogeneous enzyme immunoassay reagent and preparation method thereof

A technology of hydroxypropionase and homogeneous enzyme immunity, which is applied in the preparation of carboxylic acid esters, carboxylic acid esters/lactones, organic compounds, etc., can solve problems such as complicated operations, and achieve high immunogenicity and easy Realize and promote the effect of high accuracy

Active Publication Date: 2021-05-04
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

[0007] At present, the commonly used methods for detecting HPHPA are gas chromatography-mass spectrometry (GC-MS) and enzyme-linked immunoassay (ELISA), etc., but these methods are complicated to operate and have certain limitations in large-scale clinical application.
At present, there is a lack of HPHPA detection reagents with good stability, high sensitivity and strong specificity in the market, especially high-throughput automated detection reagents with good quality

Method used

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  • 3-(3-hydroxyphenyl)-3-hydroxypropionic acid derivative, homogeneous enzyme immunoassay reagent and preparation method thereof
  • 3-(3-hydroxyphenyl)-3-hydroxypropionic acid derivative, homogeneous enzyme immunoassay reagent and preparation method thereof
  • 3-(3-hydroxyphenyl)-3-hydroxypropionic acid derivative, homogeneous enzyme immunoassay reagent and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0078]Example 1. Synthesis of 3- (3-hydroxyphenyl) -3-hydroxypropionic acid derivative

[0079]The synthesis route of 3- (3-hydroxyphenyl) -3-hydroxypropionic acid derivative is as follows:

[0080]

[0081]The preparation process of 3- (3-hydroxyphenyl) -3-hydroxypropionic acid derivative is as follows:

[0082](1) Compound 3 synthesis:

[0083]20 g of compound 1, i.e., 3- (3-hydroxyphenyl) -3-hydroxypropionic acid (purchased from Sigma) and 33.2 g of Compound 2 (purchased from Sigma), co-dissolved in 300 ml DMF, then Join 36.4 g of K2CO3After the reaction mixing solution was prepared, the reaction mixed solution was stirred at room temperature for 12 hours, and 200 ml of purified water was added to the reaction solution, and then this solution was extracted with 300 ml of DCM, and the extraction step was repeated 3 times, Extractable organic phase through NA2SO4The drying treatment was carried out, and the residue obtained after concentration was purified by the silica gel drying column (PE: EA ...

Embodiment 2

[0086]Example 2. Synthesis of 3- (3-hydroxyphenyl) -3-hydroxypropionic acid immunogen

[0087]The 3- (3-hydroxyphenyl) -3-hydroxypropionic acid immunization in this embodiment is formed by 3- (3-hydroxyphenyl) -3-hydroxypropionic acid derivatives obtained from the BSA. The structural formula is as follows (II):

[0088]

[0089]Where the carrier is BSA.

[0090]The synthesis method of the 3- (3-hydroxyphenyl) -3-hydroxypropionic immunogen is as follows:

[0091](1) Weigh 2.72 g of dihydrogen phosphate, 4.26 g of sodium hydrogen phosphate, 8.5 g of sodium chloride, 0.95 g of magnesium chloride, and dissolve in 1 L of deionized water, adjusting pH to 8.2, making buffer A;

[0092](2) Those 3 ml of BSA, -4 ° C were weeded in 3 ml of buffer A, and a 1 mg / ml vector solution was made;

[0093](3) Weigh 3- (3-hydroxyphenyl) -3-hydroxypropionic acid derivative obtained by 3 mg Example 1, dissolved in 300 μl of buffer A at -4 ° C, 3- for 10 mg / ml of 3- (3-hydroxyphenyl) -3-hydroxypropionic acid derivative so...

Embodiment 3

[0096]Example 3. Synthesis of 3- (3-hydroxyphenyl) -3-hydroxypropionic acid immunogen

[0097]The 3- (3-hydroxyphenyl) -3-hydroxypropionic acid immunization in this example is 3- (3-hydroxyphenyl) -3-hydroxypropionic acid derivatives obtained from the oval protein. Cheng, its structural formula is as follows (II)

[0098]

[0099]Where the carrier is a folin protein.

[0100]The synthesis method of the 3- (3-hydroxyphenyl) -3-hydroxypropionic immunogen is as follows:

[0101](1) Weigh 2.3 g of pelvic phosphate, 3.8 g of sodium hydrogen phosphate, 8.0 g of sodium chloride, 0.7 g of magnesium chloride, and dissolve in 0.9L deionized water, adjust the pH to 8.0, and make buffer A;

[0102](2) Weigh 6 mg of ovantin, dissolved in 3 ml of buffer A at -4 ° C, and formulated a 2 mg / ml vector solution;

[0103](3) Weigh 2.4 mg Example 1 obtained from 3- (3-hydroxyphenyl) -3-hydroxyacionic acid derivatives, -4 ° C dissolved in 300 μL of the buffer A, 300 mg / ml 3 - (3-hydroxyphenyl) -3-hydroxypropionic acid de...

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Abstract

The invention discloses a derivative of 3-(3-hydroxyphenyl)-3-hydroxypropionic acid (HPHPA), a homogeneous enzyme immunoassay reagent and a preparation method thereof, and relates to the technical field of biological detection. Using the HPHPA derivative to prepare a highly immunogenic HPHPA immunogen and its antibody, the antibody has strong specificity and high titer, and has no cross-reaction with 92 common interfering substances. The detection reagent can realize high-throughput and rapid detection of HPPA on an automatic biochemical analyzer, with high detection accuracy, strong specificity, and significantly improved detection efficiency.

Description

Technical field[0001]The present invention relates to the field of biological detection, and more particularly to derivatives of 3- (3-hydroxyphenyl) -3-hydroxypropionic acid, homozyme immunoassay reagent and preparation method thereof, more specific to one 3- (3- Derivatives, immunogen, antibodies, enzyme coupling, detecting reagents, and preparation methods thereof.Background technique[0002]3- (3-hydroxyphenyl) -3-hydroxypropionic acid (3- (3-Hydroxyphenyl) -3-HydroxyPropionicacid, HPHPA), is shown in formula (IV):[0003][0004]HPHPA is a metabolite of 3-hydroxyphenylalanine (tyrosine analog), also a metabolite of clossed Bacillus, is discharged through urine. It is capable of causing autism-reactive autism-reactive autism-reactive autism-reactive autism.[0005]Autism Spectrum Disorders, ASDs also referred to as wide range of developmental disorders or autism, is defined as developmental disorders with social interaction and communication barriers, scope of interest and repeated sten...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C59/68C07C51/09C07C69/712C07C67/31C07K14/765C07K14/77C07K14/795C07K14/47C07K16/44
Inventor 虞留明王益超娄明星
Owner 苏州博源医疗科技有限公司
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