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Ligand conjugated quantum dot nanoparticles and methods of detecting DNA methylation using same

A nanoparticle, methylation technology, applied in biochemical equipment and methods, nanotechnology for sensing, nanotechnology, etc., can solve problems such as detection and limitation of difficult to achieve abnormal DNA methylation

Inactive Publication Date: 2019-04-26
NANOCO TECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a result, the detection of aberrant DNA methylation is difficult to achieve, which limits the understanding of the role and patterns of DNA methylation in specific diseases, such as cancer

Method used

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  • Ligand conjugated quantum dot nanoparticles and methods of detecting DNA methylation using same
  • Ligand conjugated quantum dot nanoparticles and methods of detecting DNA methylation using same
  • Ligand conjugated quantum dot nanoparticles and methods of detecting DNA methylation using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Synthesis of Nontoxic Quantum Dots

[0085] A molecular seeding process is used to generate non-toxic quantum dots (QDs). Briefly, the preparation of non-functionalized indium-based quantum dots emitting in the 500-700 nm range was carried out as follows: dibutyl ester (about 100 ml) and myristic acid (MA) (10.06 g) were placed in a three-necked flask, Vacuum degassed at about 70°C for 1 hour. Thereafter, nitrogen gas was introduced and the temperature was raised to about 90°C. Add about 4.7g of ZnS molecular clusters [Et 3 NH] 4 [Zn 10 S 4 (SPh) 16 ], and the mixture was stirred for about 45 minutes. The temperature was then raised to about 100°C, followed by the dropwise addition of In(MA) 3 (1M, 15ml), followed by the addition of trimethylsilylphosphine (TMS) 3 P (1M, 15ml). The reaction mixture was stirred while raising the temperature to about 140°C. At 140°C, indium myristate (In(MA)) dissolved in di-n-butyl sebacate was further added dropwise. 3 ) (1M...

Embodiment 2

[0089] Water-soluble surface modified QD

[0090] Provided herein is one embodiment of a method for producing and using melamine hexamethoxymethylmelamine (HMMM) modified fluorescent nanoparticles as drug delivery vehicles. The unique melamine-based coating has excellent biocompatibility, low toxicity and very low non-specific binding. These unique features allow for a wide range of biomedical applications in vitro and in vivo.

[0091] An example of the preparation of suitable water-soluble nanoparticles is provided as follows: 200 mg of cadmium-free quantum dot nanoparticles emitting red at 608 nm as core material, an alloy containing indium and phosphorous is dispersed in In toluene (1 ml) containing isopropyl myristate (100 µl). Contains isopropyl myristate as a ligand interactor. The mixture was heated at 50°C for about 1-2 minutes, then slowly shaken at room temperature for 15 hours. A mixture containing hexamethoxymethylmelamine (HMMM) (CYMEL 303, purchased from Cyt...

Embodiment 3

[0104] Water-soluble QDs including targeting ligands

[0105] In certain embodiments, water-soluble QDs are modified to include targeting ligands added to the QDs. Therefore, in one embodiment, quantum dot nanoparticles are synthesized, which are non-toxic and water-soluble (biocompatible) and whose surface is equipped with functionalities capable of coupling (COOH, OH, NH 2 , SH, azides, alkynes). Due to functional groups that can be added to QDs, such as the COOH functional group provided in Example 2 herein, QDs can be modified to include targeting ligands that allow QDs to selectively identify methylation in samples, cells, and tissues DNA. Targeting ligand-modified QDs are illuminated and emit light for detection.

[0106] In an exemplary embodiment, the water-soluble nontoxic QDs are or become carboxy-functionalized. COOH-QDs are attached to methylated DNA using chemical methods such as carbodiimide ligation using water-soluble 1-ethyl-3-(-3-dimethylaminopropyl)carbo...

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Abstract

The present disclosure relates to conjugated quantum dot nanoparticles, to methods of making such conjugated quantum dot nanoparticles, and to methods of detecting DNA methylation using such conjugated quantum dot nanoparticles. In one embodiment, the quantum dot is conjugated to an antibody that recognizes methylated DNA bases, such as an anti-5-methylcytosine antibody, an anti-5- hydroxymethylcytosine antibody, an anti-5-formylcytosine antibody, an anti-5-carboxylcytosine antibody or an anti-N6-methyladenine antibody.

Description

technical field [0001] Embodiments disclosed herein relate to quantum dot nanoparticles coupled to ligands, particularly quantum dot nanoparticles coupled to methylation-specific binding ligands. Embodiments also include methods of making such coupled quantum dot nanoparticles, and methods of using such coupled quantum dot nanoparticles to detect DNA methylation. Background technique [0002] DNA methylation is an epigenetic process used by cells to control gene expression and has important functions in normal and disease biology. Alteration of DNA methylation by, for example, aging and disease can lead to irregular methylation, which can lead to many complex diseases in mammals, such as heart disease, diabetes, neurological diseases, and cancer. See, eg, Okhov-Mitsel et al., Cancer Medicine, 237-260, 2012 Review. For example, DNA methylation contributes to carcinogenesis by silencing tumor suppressor genes. See, eg, Jones et al., Nat. Rev. Genet., 3, 415-428, 2002. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00G01N33/53G01N33/58B82Y15/00
CPCA61K49/0058A61K49/0067G01N33/5308B82Y15/00G01N33/00G01N33/588C12Q1/6804G01N33/54346G01N33/54353C12Q1/00G01N33/533G01N33/582
Inventor 伊马德·纳萨尼
Owner NANOCO TECH LTD
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