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Quantum dot nanocomposite and preparation method and application thereof

A technology of nanocomposites and quantum dots, applied in the direction of nanotechnology, nanooptics, nanotechnology, etc., to achieve the effect of facilitating the path

Pending Publication Date: 2018-11-23
FIRST AFFILIATED HOSPITAL OF DALIAN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Labeling exosomes to explore their metabolism in the body has become a major challenge

Method used

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  • Quantum dot nanocomposite and preparation method and application thereof
  • Quantum dot nanocomposite and preparation method and application thereof
  • Quantum dot nanocomposite and preparation method and application thereof

Examples

Experimental program
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Embodiment 1

[0028] Quantum dot aptamer fluorescent probe preparation:

[0029] The water-soluble quantum dots with amino groups were passed through the cross-linking agent sulfo-SMCC [(N-maleimidomethyl) cyclohexane-1-carboxylic acid succinimide ester, sodium salt] Sulfo- SMCC was coupled to a CD63 aptamer with the following thiol-modified sequence: CACCCCACCTCGCTCCCGTGACACTAATGCTA.

[0030] The specific coupling steps are as follows:

[0031] (1) Thaw Sulfo-SMCC and CD63 aptamer before the experiment.

[0032] (2) Add 88 μL (8 μM) of amino water-soluble quantum dots into the reactor under dark conditions, add an equal volume of 4 μM Sulfo-SMCC, and react on a rapid shaking table at room temperature for 1 h under dark conditions.

[0033] (3) The above reaction solution was centrifuged at 10,000 rpm for 5 min to remove the precipitate.

[0034] (4) Add the supernatant to the desalting column NAP-5columns, elute with 10mL PBS, collect the fluorescent part, about 800μL.

[0035] (5) Add...

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Abstract

The invention provides a quantum dot nanocomposite, and a preparation method and application thereof. Specifically, the quantum dot nanocomposite is formed in the way that a biological recognition factor CD63 aptamer of an exosome membrane is chemically coupled on the surface of a quantum dot probe. The quantum dot nanocomposite is applied to tracing the interactions of an exosome sourced from a stem cell and cells of metabolic organs (liver, lung and kidney) by using a confocal microscope imaging technology. The specific condition is that the ratio of Apt to QDs is 2 muL to 1 mug. The co-incubation time of the exosome sourced from the stem cell and the cells of the metabolic organ cells is 5-8 hours, and the imaging ratio of the exosome to the cells is (20-40) mug to (2-4)*10<4> / mL. The exosome is used as a technology platform, the metabolic pathway of the exosome marked by the quantum dot-aptamer nanoprobe is researched in vitro, and technical support is provided for in-vivo and clinical research such as subsequent exosome drug load.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a quantum dot nanocomposite, its preparation method and application. Background technique [0002] Exosomes (Exosomes, Exo) are vesicles with a diameter of 30-150 nm secreted by cells, which undertake functions such as intercellular communication and material transport. Because exosomes carry proteins and RNA, etc., and have low immunogenicity, their use as disease markers and targeted drug carriers has become a hotspot. Studies have shown that stem cell-derived exosomes have a repair function, especially in degenerative and neuroinflammatory diseases. In addition, specific markers associated with exosomes can be used to monitor cancer. In order to further use stem cell-derived exosomes for clinical treatment, researchers need to fully understand their metabolism. The research on exosomes is still in the early stage, and the metabolic pathways of exosomes secreted by d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/06B82Y20/00B82Y30/00B82Y40/00A61K49/00
CPCA61K49/0067C09K11/06B82Y20/00B82Y30/00B82Y40/00
Inventor 刘晶沈丽明马静云
Owner FIRST AFFILIATED HOSPITAL OF DALIAN MEDICAL UNIV