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Screening methods and kits for DNA-encoded molecular libraries

A screening method and molecular library technology, applied in the field of DNA-encoded molecular library screening methods and kits, can solve problems such as the influence of target activity, and achieve the effects of improving efficiency, increasing concentration, and expanding the screening range

Active Publication Date: 2020-12-01
SHENZHEN NEWDEL BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In some methods, a DNA fragment with a cross-linking group is first used to mark the target, and then the DNA fragment is used to complement the DNA molecules in the DNA-encoded molecular library to realize the screening of compound molecules. However, since this method uses covalent The target is marked by cross-linking, and strong denaturing conditions are required for subsequent elution, which has a great impact on the activity of the target on living cells

Method used

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  • Screening methods and kits for DNA-encoded molecular libraries
  • Screening methods and kits for DNA-encoded molecular libraries
  • Screening methods and kits for DNA-encoded molecular libraries

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] In this example, the specific protein labeling of living cell membrane proteins is verified, and the specific process is as follows:

[0075] (1) Preparation of MDA-MB-435S cell-integrin-RGDfK cyclic peptide-A chain-FAM complex

[0076] MDA-MB-435S cells were used as the research system, and the integrin protein on the surface of MDA-MB-435S cells was used as the research target protein;

[0077] Using the RGDfK cyclic peptide as a small molecule ligand that specifically binds to integrin, the RGDfK cyclic peptide is connected to the first DNA through a cleavable group to obtain the RGDfK cyclic peptide-A chain complex, which is labeled as the A chain;

[0078] In order to visually show that the A chain binds specifically to the target, a fluorescent group fluorescein (FAM) was connected to the end of the first DNA of the A chain away from the RGDfK cyclic peptide to obtain an A chain-FAM complex.

[0079] Such as figure 1 As shown, the A chain-FAM complex was incubat...

Embodiment 2

[0088] In this example, the DNA-encoded molecular library is screened for cell surface integrins. The specific process is as follows: Figure 5 Shown:

[0089] A library of DNA-encoded molecules is provided. The DNA-encoded molecule library contains 30 million encoded compounds, each encoded compound includes sequentially linked compounds, a cleavable group, a second DNA (marked as n1) and a compound-specific encoded DNA ( Marked as n2), at the same time, the DNA encoding molecule library also contains an encoding RGDfK cyclic peptide capable of specifically recognizing integrin protein, as a positive control;

[0090] After fully mixing and incubating the A chain with the MDA-MB-435S cells, the small molecule ligand RGDfK cyclic peptide of the integrin protein connected in the A chain recognizes the integrin protein bound to the surface of the MDA-MB-435S cells, and obtains the MDA-MB-435S cell surface-containing integrin protein Mixed system of MB-435S cell-integrin-A chain...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a screening method of a DNA encoded molecule library. The method comprises the following steps: providing a first compound and a target spot, wherein the first compound comprises, sequentially connected, a first DNA, a cleavable group and a ligand capable of specifically binding to the target spot; incubating thefirst compound and the target spot to obtain a second compound; providing a DNA encoded molecule library, wherein an encoded compound in the DNA encoded molecule library contains a second DNA, and a complementary base sequence exists between the second DNA and the first DNA; incubating the DNA encoded molecule library and the second compound to obtain a first mixed system; providing a cut-off reagent, incubating the cut-off reagent and the first mixed system, and carrying out separating to obtain a second mixed system; and carrying out denaturation dissociation on the second mixed system to obtain a dissociation product, analyzing DNA sequence information in the dissociation product, and determining the target encoded compound according to the DNA sequence information.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a screening method and kit for a DNA-encoded molecular library. Background technique [0002] In contemporary drug research and development, high-throughput and large-scale screening is an indispensable means for new drug research and development by constructing a large candidate drug molecular library for disease drug targets. Major pharmaceutical companies in the world today have large-scale molecular libraries and large-scale screening platforms for new drug research and development. However, traditional molecular libraries and screening platforms are costly, have high technical barriers, and are complex in management and operation, which severely restricts high-throughput Screening development and application issues. [0003] In recent years, DNA-encoded molecular library technology has gradually developed and has become an emerging screening method in drug developme...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C40B40/08C40B50/16C40B70/00G01N33/53
CPCC40B40/08C40B50/16C40B70/00G01N33/5308
Inventor 熊峰周海鹏
Owner SHENZHEN NEWDEL BIOTECH CO LTD
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