Method for seeking micromolecule chemical drug target spots by combining quantum dot nanometer fluorescent probe and protein chip

A nano-fluorescence probe and protein chip technology, which is applied in the field of quantum dot nano-fluorescence probes combined with protein chips to find small molecule chemical drug targets, can solve the problems of low fluorescence quantum yield, difficult detection, poor optical stability, etc., and achieve The effect of broad application prospects

Inactive Publication Date: 2012-07-04
CGENETECH (SUZHOU CHINA) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this fluorescent dye has major defects, such as low fluorescence quantum yield; poor optical stability; strict requirements on the optical part of the detection system; large overlap between excitation spectrum and emission, which brings difficulties to detection; Probes are excited with the same excitation light
[0005]

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Add 300 μl of ethyl orthosilicate containing 27% (v / v) absolute ethanol to 50 ml of CdSe (590) (self-made) (590 refers to the peak position of the emission spectrum of CdSe at 590 nm) nanoparticle aqueous solution with a concentration of about 10 nM. The mixed solution of ester and 20 μl ammonia water, after reacting for 2-3 days, obtain the solution that is still clear and transparent, get some samples after freeze-drying and analyze through X-ray diffraction (XRD) and X-ray energy spectrum (XPS), the result shows that CdSe It shows that silicon dioxide has been coated, that is, CdSe / SiO2 has been formed. (When preparing CdSe / SiO2, the amount of ethyl orthosilicate, absolute ethanol and ammonia should be controlled, otherwise particle aggregation will be caused).

[0020] Add CdSe / SiO2 to the solution of absolute ethanol and silane coupling agent H2NCH5Si(OC2H5)3 (in the reaction system, CdSe / KH550-10: 1.5, absolute ethanol / water=10), and react at 60°C for 2h to obtain...

Embodiment 2

[0022] Dissolve small molecule chemicals in PBS buffer solution of pH 7, add 1-ethyl-3(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), react at room temperature for 1h, then add indicated The aqueous solution of quantum dot CdSe / ZnS(590) (self-made) nanoparticles modified with mercaptoethylamine was reacted overnight at room temperature, and the quantum dots were connected to the drug through the covalent reaction of amino and carboxyl groups. In the reaction solution, CdSe / ZnS is about 50nM, and the molar ratio of CdSe / ZnS, small molecule chemical drug and EDC is listed as 1:100:120. The free small molecule chemical drug and EDC are separated by gel column to obtain " CdSe / ZnS-Small Molecule Chemicals" ligation product. The product emits the same fluorescence as CdSe / ZnS, that is, red fluorescence, when excited by light at a wavelength of 488mm. HPLC experiments show that the retention time of "CdSe / ZnS-small molecule chemical drug" is about 1 min slower, indicating ...

Embodiment 3

[0024] Co-incubation of "quantum dots-small molecule chemical drugs" nanoprobes with protein chips: the protein chip in the kit is used in this work. Although this chip is not specially designed for this experiment, a variety of specific antigens are immobilized on its surface. They are expressed on the surface of normal cells and tumor cells, especially on the surface of tumor cells. The experiment was operated according to the method of the kit. The "quantum dot-small molecule chemical drug" nanoprobe solution and the chip were incubated at 37°C for 30-40min, washed, and analyzed by laser confocal scanning. It was found that there were a small number of fluorescent spots on the chip, indicating that small molecule chemicals may act on a variety of tumor cell surface antigens.

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Abstract

The invention discloses a method for seeking micromolecule chemical drug target spots by combining a quantum dot nanometer fluorescent probe and a protein chip, comprising the steps of: connecting a quantum dot with a micromolecule chemical drug to form a quantum dot-micromolecule chemical drug fluorescent probe; then directly incubating the fluorescent probe and the protein chip together; after washing, determining the binding sites (target spots) of specificities between the drug and known proteins fixed on the chip according to fluorescence given out by the optical excitation of the quantum dot on the protein chip; and analyzing the proteins at the binding sites through computer software. According to the method for seeking micromolecule chemical drug target spots by combining the quantum dot nanometer fluorescent probe and the protein chip, the micromolecule chemical drug and the target proteins having efficacy on cells can be simply, conveniently and rapidly sought, and an efficient technical means for screening novel micromolecule chemical drugs in a high-throughput manner can be provided.

Description

technical field [0001] The invention provides a method for finding the action target of small molecule chemical drugs by combining quantum dot nanotechnology with protein chip technology, which belongs to the intersecting fields of materials, chemical synthesis, chemical industry, pharmacy and biology, and specifically relates to a method of using nanotechnology and protein chip Combining technologies to find the target of small molecule chemical drugs, especially the method of finding the target protein of small molecule chemical drugs on the protein chip through the fluorescence of quantum dot nanoparticles. Background technique [0002] Over the years, people's studies have shown that most drugs achieve their desired drug effects by interfering with the functions of enzymes or receptors in cells. Enzymes or receptors are proteins, and there are many small molecules on their surfaces that can be occupied. Vacancy. When a small molecule binds to a vacancy on an enzyme or re...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543C09K11/06
Inventor 余强
Owner CGENETECH (SUZHOU CHINA) CO LTD
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