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T cell and nanoparticle connection method based on click chemistry

A nanoparticle and connection method technology, applied in the field of nanomedicine, can solve the problems of low efficiency, unstable connection method, poor specificity, etc., and achieve the effects of avoiding false signals, fast and easy labeling, and stable labeling products

Active Publication Date: 2021-06-18
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Aiming at the defects of unstable connection method, low efficiency and poor specificity in the prior art, the present invention provides a method for connecting T cells and nanoparticles based on click chemistry

Method used

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  • T cell and nanoparticle connection method based on click chemistry
  • T cell and nanoparticle connection method based on click chemistry
  • T cell and nanoparticle connection method based on click chemistry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The working principle of the present invention using click chemistry to label cells with fluorescent quantum dots can be found in figure 1 . Through the diphenylcyclooctyne group (-DBCO)-azide (-N 3 ) linked click chemistry method, the nanoparticles (INPs-DBCO) and the azide (-N 3 ) covalently bind to form a highly efficient link between T cells and nanoparticles.

[0036] Concrete steps of the present invention are as follows:

[0037] Step 1: Preparation of T cells

[0038] (1) T cell extraction and expansion: the peripheral blood of healthy volunteers was collected, and human peripheral blood mononuclear cells (PBMCs) were isolated. The specific operation is: collect blood in the anticoagulant tube, add 1:1 normal saline, another tube is added with a separation solution equal to the volume of normal saline, the mixture of normal saline and blood is added to the separation solution along the tube wall, then 800g, 30min After centrifugation, the layers are general...

Embodiment 2

[0047] Measuring the expression of azide group metabolism on the surface of T cell membrane by flow cytometry

[0048] like figure 2 As shown by flow cytometry data, the azide group (-N 3 ) was labeled with FITC dye, and the flow peak of the T cell group incubated with the sugar molecule tetraacetyl-N-azidoacetylmannosamine (Ac4ManAz) was separated from the flow peak of the control group (Control) that was not incubated with the sugar molecule The obvious difference indicates that the T cell group incubated with sugar molecules was successfully labeled with FITC dye, and the azide group was successfully expressed on the surface of the T cell membrane.

Embodiment 3

[0050] Particle Size Detection of Indocyanine Green Polymer Nanoparticles (INPs)

[0051] like image 3 As shown, the INPs prepared by the ultrasonic hydration method were used to detect the particle size of the nanoparticles using a Malvern particle size analyzer, and the average particle size of the INPs nanoparticles was about 160nm.

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Abstract

The invention relates to the field of nano medicine, in particular to a T cell and nano particle connection method based on click chemistry. Through a click chemistry method of diphenyl cyclooctyne group-azide connection, nanoparticles and azide groups on cell membrane surface protein are covalently bound to form efficient connection of T cells and the nanoparticles. An existing connection method is unstable, low in efficiency and poor in specificity, and the T cell and nano particle connection method based on click chemistry overcomes the defects, is simple and easy to implement, mild in reaction condition and stable in connection, and can realize nano engineering transformation of the T cells.

Description

technical field [0001] The invention relates to the field of nanomedicine, in particular to a method for connecting T cells and nanoparticles based on click chemistry. Background technique [0002] Click chemistry is a modular synthesis concept first proposed in 2001 by Nobel Prize winner K.Barry Sharpless. It is a new method of combinatorial chemistry that selects readily available raw materials, realizes carbon heteroatom connection through modular, reliable, high-efficiency, and high-selectivity chemical transformations, and rapidly synthesizes various new compounds at low cost. [0003] At present, the most widely used click chemistry is the copper ion-catalyzed Huisgen dipolar cycloaddition reaction of terminal alkynes and azides. However, due to the introduction of copper ions as catalysts during the click reaction, the physiological toxicity of residual copper ions will lead to the degradation of DNA and the denaturation of proteins. The new copper-free click chemis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783C12N5/00C08G81/00
CPCC12N5/0636C12N5/0006C08G81/00Y02A50/30
Inventor 蔡林涛陈泽郑明彬潘宏尹婷马爱青邢婕华罗英梅
Owner SHENZHEN INST OF ADVANCED TECH
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