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Preparation method of high-throughput liquid crystal detection platform for screening enzyme inhibitor by inducing aptamer to release through enzyme catalysis

A detection platform and aptamer technology, which is applied in the field of analysis and detection, can solve problems such as restricting development, and achieve good effects, high sensitivity, and good selectivity

Active Publication Date: 2021-06-11
SHANDONG ANALYSIS & TEST CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The inventors found that there is currently no simple platform that can quickly and effectively screen enzyme inhibitors, which limits the development of research in this direction

Method used

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  • Preparation method of high-throughput liquid crystal detection platform for screening enzyme inhibitor by inducing aptamer to release through enzyme catalysis
  • Preparation method of high-throughput liquid crystal detection platform for screening enzyme inhibitor by inducing aptamer to release through enzyme catalysis
  • Preparation method of high-throughput liquid crystal detection platform for screening enzyme inhibitor by inducing aptamer to release through enzyme catalysis

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preparation example Construction

[0053] A second aspect of the present invention provides a method for preparing a high-throughput liquid crystal biosensor for screening enzyme inhibitors by enzymatically inducing aptamer release, comprising:

[0054] Pretreatment of glass substrates;

[0055] Add liquid crystal molecules to the chloroform solution containing surfactant (OTAB), and then remove the chloroform with nitrogen to obtain a mixture of liquid crystal and OTAB, which is added dropwise to the surface of the copper mesh;

[0056]The inhibitory drug to be tested is mixed with substrate-converting specific enzyme, small molecule substrate, and aptamer DNA successively, and added dropwise on the above-mentioned copper grid to obtain the product.

[0057] The invention adopts the copper mesh method to observe the optical phenomenon of the liquid crystal, and has high sensitivity, short time consumption and simple preparation method.

[0058] A third aspect of the present invention provides a method for hig...

Embodiment 1

[0110] A high-throughput liquid crystal detection platform for screening xanthinease inhibitors using enzyme-catalyzed aptamer release to detect the enzyme inhibition efficiency of febuxin

[0111] The liquid crystal sensor used was prepared as follows:

[0112] a. Soak the glass sheet in the washing solution at 75°C for 30 minutes, then wash it with a large amount of ultrapure water, methanol, and ethanol, blow it dry with nitrogen, and dry it in an oven at 120°C for 10 to 20 hours.

[0113] b. At room temperature, soak the glass sheet in DMOAP with a volume fraction of 1% for 20 minutes, then wash it with ultrapure water, and dry it with nitrogen to obtain a glass substrate.

[0114] Several copper meshes were pipetted onto the resulting glass substrates.

[0115] Dissolve OTAB in chloroform to make a chloroform solution with a concentration of 1 mMOTAB.

[0116]c. Mix 10 μL of 5CB molecules with 90 μL of 1 mM OTAB solution, vortex for 30 seconds to obtain a final concentr...

Embodiment 2

[0124] Example 2: Synthesis of various xanthinease inhibitors and screening of effective drugs using a high-throughput liquid crystal detection platform for screening xanthinease inhibitors by inducing aptamer release through enzyme catalysis

[0125] The preparation of liquid crystal sensor is identical with embodiment 1

[0126] S1. Pre-mix the compound to be detected with xanthine oxidase and incubate for 30 minutes; then incubate with xanthine for 30 minutes, and then incubate with xanthine aptamer DNA for 60 minutes.

[0127] S2. Pipette 40 μL of the prepared mixture onto a liquid crystal sensor, and observe using a polarizing microscope.

[0128] The synthesized xanthine enzyme inhibitory drugs are all benzotriazole compounds and their derivatives. Through the above-mentioned liquid crystal detection platform, three suitable enzyme inhibitor drugs can be screened out with high throughput and effectively. Each structure is shown in Figure 5.

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Abstract

The invention belongs to the field of analysis and detection, and relates to a preparation method of a high-throughput liquid crystal detection platform for screening an enzyme inhibitor by inducing aptamer release through enzyme catalysis. The high-throughput liquid crystal detection platform comprises the following components of: a glass slide, a copper net, liquid crystal molecules, a surfactant, aptamer coding DNA, a substrate and a substrate specific invertase. The components are utilized to modify a liquid crystal biosensor, efficient screening of the enzyme inhibitor is achieved, and the effect of the platform is better than that of an existing screening method at present. The detection method provided by the invention has the advantages of high sensitivity and short time consumption; and the preparation method has the advantages of simplicity, good selectivity, rapidness, simplicity, low cost and no need of large-scale instruments, and has a good application prospect in high-throughput screening of enzyme inhibitors.

Description

technical field [0001] The invention belongs to the technical field of analysis and detection, and in particular relates to a liquid crystal biosensor for screening enzyme inhibitors and high-throughput screening for enzyme inhibitors. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily to be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] An enzyme inhibitor is a substance that can inhibit the activity of a specific enzyme related to a certain disease in an organism, thereby obtaining a curative effect. Enzyme inhibitors are substances that specifically act on certain groups of enzymes to reduce the activity of the enzyme or even completely inactivate the enzyme to reduce the rate of the enzymatic reaction. At present...

Claims

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Application Information

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IPC IPC(8): G01N21/84
CPCG01N21/84
Inventor 胡琼政武文丽夏爽
Owner SHANDONG ANALYSIS & TEST CENT
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