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Method of labeling exosome, exosome labeled by AIE fluorescent molecular marker, and application thereof

A technology of fluorescent molecules and exosomes, which can be used in preparations for in vivo experiments, pharmaceutical formulations, bone/connective tissue cells, etc., and can solve problems such as high quenching coefficient, easy quenching, and errors.

Active Publication Date: 2019-03-01
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the most commonly used fluorescent dyes have high quenching coefficients, resulting in short half-lives and easy quenching, and they can form dye aggregates in aqueous solutions similar to exosomes, leading to uptake in exosomes. error in experiment

Method used

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  • Method of labeling exosome, exosome labeled by AIE fluorescent molecular marker, and application thereof
  • Method of labeling exosome, exosome labeled by AIE fluorescent molecular marker, and application thereof
  • Method of labeling exosome, exosome labeled by AIE fluorescent molecular marker, and application thereof

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Embodiment

[0050] This example is used to illustrate the method for labeling exosomes of the present disclosure.

[0051] Take 0.8 μL of 5 mmol / L AIE fluorescent molecule, add it to the extracted 100 μl exosome sample containing 100 μg protein, make up to 1000 μL with PBS, shake and mix well, and perform fluorescent labeling in a dark room at room temperature 37°C for 2 hours. Then the above mixture was transferred into an ultracentrifuge tube, filled with PBS, centrifuged at 100,000 rcf at 4°C for 60 min, and then the supernatant was removed to obtain AIE fluorescent molecularly labeled exosomes.

[0052] The exosomes labeled with AIE fluorescent molecules were resuspended in 50 μL PBS, aliquoted and stored in a -80°C refrigerator for later use.

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Abstract

The disclosure relates to a method of labeling an exosome, the exosome labeled by an AIE fluorescent molecular marker, and an application thereof. The method includes: mixing a labeling solution containing an AIE fluorescent molecule and the exosome for fluorescent labeling. In the disclosure, a specific aggregation-induced emission fluorescent molecule can stably and high-effectively label the exosome without change on the structure and component of the exosome and influence on bio-functions of same. The labeled exosome can be used for preparing a tracing medicine, which can help further researching on functions of the exosome.

Description

technical field [0001] The present disclosure relates to the field of biotechnology, in particular, to a method for labeling exosomes, AIE fluorescent molecule-labeled exosomes and applications thereof. Background technique [0002] Exosomes are nanoscale extracellular vesicles with a double-membrane structure and a diameter of about 30-100 nm secreted by living cells. They naturally exist in various body fluids such as blood, saliva, urine, and mother fluid. As an important part of extracellular vesicles, exosomes have become an important form of intercellular communication, and the intercellular communication mediated by them can provide neighboring cells with a large number of biologically active signal molecules, including DNA, RNA, microRNA and proteins Wait. Exosomes released outside cells can enter target cells through different pathways to exert their biological functions. First, exosomes can trigger signal transduction pathways in target cells by binding to recept...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00C12N5/0775
CPCA61K49/0021A61K49/0097C12N5/0668
Inventor 王悦冰曹红梅岳志伟丁丹李宗金
Owner NANKAI UNIV