Method for detecting receptor-ligand binding blocking capacity of compound by using HTRF technology

A technology of technical detection and combination ability, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of difficult popularization and high price, and achieve the effects of stable signal, saving experimental time, and long-lasting fluorescence

Pending Publication Date: 2021-11-09
SUZHOU GENHOUSE PHARMA RES & DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0014] To sum up, the existing detection methods all require special instruments and are expensive, and some even require custom-made consumables, such as isotopic markers, which are difficult to popularize

Method used

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  • Method for detecting receptor-ligand binding blocking capacity of compound by using HTRF technology
  • Method for detecting receptor-ligand binding blocking capacity of compound by using HTRF technology

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0039] Step 1, the androgen receptor containing the GST tag is labeled with the GST antibody of the chimeric lanthanide Tb;

[0040] Step 2, labeling androgen with FITC;

[0041] Step 3, incubating the serially diluted Darolutamide together with the labeled androgen receptor and androgen at room temperature;

[0042] Step 4, detecting the receptor fluorescence signal, and calculating the ability of the compound to block receptor-ligand binding;

[0043] Such as figure 2 As shown, Darolutamide dose-dependently blocks androgen receptor and androgen binding.

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Abstract

The invention discloses a method for detecting the receptor-ligand binding blocking capacity of a compound by using an HTRF technology, and belongs to the field of biochemical detection. The method for detecting the receptor-ligand binding capacity by utilizing the HTRF technology comprises the following steps of: 1, carrying out fluorescent donor labeling on a ligand; 2, carrying out fluorescent receptor labeling on a receptor; 3, co-incubating the receptor, the ligand and a tested compound; and 4, detecting a fluorescence signal of the receptor, and calculating the receptor-ligand binding blocking capability of the compound. The method has the advantages of homogeneous reaction, simple operation and high throughput test.

Description

technical field [0001] The invention belongs to the field of high-throughput compound screening, and specifically relates to a method for detecting compound blocking receptor-ligand binding ability by using HTRF technology. Background technique [0002] Receptor is a cellular protein component formed during the evolution of cells, which can recognize a certain trace chemical substance in the surrounding environment, first bind to it, and trigger subsequent physiological responses through the intermediary information transduction and amplification system or pharmacological effects. [0003] Since Langley put forward the receptor theory 100 years ago, receptors have been proved to be objectively existing entities with various types and mechanisms of action have been elucidated. Now receptors are no longer an empty and general concept. [0004] The content of receptor molecules in cells is very small, and 1 mg of tissue generally only contains about 10 fmol. A substance that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/58G01N33/53
CPCG01N21/6408G01N33/582G01N33/53
Inventor 张涛李家鹏王奎锋
Owner SUZHOU GENHOUSE PHARMA RES & DEV CO LTD
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